Oxford Laboratory Technology

Isolation of Casein from Milk

2011-12-25T03:04:00.001-08:00

Isolation and Identification of Casein From Milk Course Notes

Milk is the probably the most nutritionally complete food found in nature. Whole milk contains vitamins (principally thiamine,riboflavin, panthothenic acid and vitamins A, B₁₂ and D), minerals (calcium, sodium, phosphorus, potassium, and trace minerals), proteins (which include all the essential amino acids), carbohydrates (mostly lactose), and lipids (fats). Whole milk is an oil in water emulsion, containing approximately 4% fat dispersed as very small (micron sized) globules. The fat emulsion is stabilized by complex phospholipids and proteins that are absorbed on the surface of the emulsion. Since the fat in milk is so finely dispersed it is more easily digested than fats from any other source.

Read Full Protocol Here

Isolation of Casein, Lactose, and Albumin from Milk

Milk is a food of exceptional interest. Not only is milk an excellent food for the very young, but humans

have also adapted milk, specifically cow’s milk, as a food substance for persons of all ages. Many

specialized milk products like cheese, yogurt, butter, and ice cream are staples of our diet.

Read Full Protocol Here.

Experiment 11: Isolation and Characterization of Casein from Milk

Adapted from Experiment 21, “Isolation of Protein, Carbohydrate and Fat from Milk”, in

Mohr. S.C., Griffin, S.F., and Gensler, W. J. Laboratory Manual for Fundamentals of

Organic and Biological Chemistry by John McMurry and Mary E. Castellion,: nglewood

Cliffs, Prentice-Hall, 1994 and Wayne P. Anderson (4/2002)

Read Full Experimental Protocol Here

ISOLATION OF CASEIN FROM MILK

Purpose: In this lab you will be isolating the proteins casein and lactalbumin from a sample of milk. You

will use these values to determine the percent protein in milk compared to the listed value on the box.

Read Full Experimental Protocol here.

Polony PCR Amplification

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This section outlines the protocol for single-molecule amplification within the acrylamide gel. The protocol given here may differ in specifics from what is presented here, but reflects what we are doing currently (Summer 2003). We have tried to include excessive detail here but let us know if anything is unclear. The general steps are:

1.    Cast acrylamide gels
2.    Diffuse in PCR reagents
3.    PCR amplification
4.    Slide clean-up

Further Protocol

Long PCR Protocol Reagents and Guidelines

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General Guidelines for Long PCR Conditions and Enzyme Mixtures
Efficient Long PCR results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofreading polymerase (3' to 5' exo) is present at a lower concentration. Following the results of Cheng et al. (1) we have had success using Tth (ABI/Perkin-Elmer) as the main-component polymerase and Vent (New England Biolabs) as the fractional-component polymerase. Other combinations of proofreading and non-proofreading polymerases have been used successfully for many applications. The buffer listed below works well with Tth and Vent, but not with others. If you are interested in using other polymerases make sure that you use compatible buffers. Error rates for other polymerases can be found at http://research.nwfsc.noaa.gov/protocols/taq-errors.html

Read Full Protocol Here

Qiagen RNeasy Plant RNA Isolation

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RNAisolation

RNeasy Kitsare designed to isolate total RNA from small quantities of starting material. Theyprovide a fast and simple method for the preparation of up to 100 µg total RNAfrom animal cells and tissues, bacteria, and yeast (RNeasy MiniKits)or plant cells and tissues and filamentous fungi (RNeasy PlantMini Kits). RNeasy Plant Mini Kits are fast and avoid tedious methods, suchas CsCl step-gradient ultracentrifugation and alcohol precipitation steps, ormethods involving the use of toxic substances such as phenol and/or chloroform.The purified RNA is ready for use in standard downstream applications such asRT-PCR, Northern, dot, and slot blotting, poly A+ RNA selection, primerextension, RNase and S1 nuclease protection, cDNA synthesis, differentialdisplay, expression-array and expression-chip analysis.

Further Protocol Information

Formaldehyde Agarose Gel Electrophoresis Protocol

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The followingprotocol for formaldehyde-agarose gel electrophoresis gives enhancedsensitivity for gel and subsequent analysis (e.g. northern blotting). A keyfeature is the concentrated RNA loading buffer that allows a larger volume ofRNA sample to be loaded onto the gel than conventional protocols (e.g. Sambrook,J. et al., eds. (1989) Molecular cloning — a laboratorymanual, 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor LaboratoryPress).

1.2% FormaldehydeAgarose gel preparation

To prepare FormaldehydeAgarose gel (1.2% agarose) of size 10 x 14 x 0.7 cm, mix:

1.2 g agarose

10 ml 10x FormaldehydeAgarose gel buffer (see composition below)

Add RNase-freewater to 100 ml

If smaller orlarger gels are needed, adjust the quantities of components proportionately. Heatthe mixture to melt agarose. Cool to 65°C in a water bath. Add 1.8 ml of 37% (12.3M) formaldehyde (toxic) and 1 µl of a 10 mg/ml ethidiumbromide (mutagenic) stock solution.Mix thoroughly and pour onto gel support. Prior to running the gel, equilibratein 1x Formaldehyde Agarose gel running buffer for at least 30 min.

RNA samplepreparation for Formaldehyde Agarose gel electrophoresis

Add 1 volumeof 5x loading buffer per 4 volumes of RNA sample (for example 10 µl of loadingbuffer and 40 µl of RNA) and mix.

Incubate for3–5 min at 65°C, chill on ice, and load onto the equilibrated FormaldehydeAgarose gel.

Gel runningconditions

Run gel at 5–7V/cm in 1x Formaldehyde Agarose gel running buffer.

Compositionof Formaldehyde Agarose gel buffers

10x FormaldehydeAgarose Gel buffer

200 mM3-[N-morpholino]propanesulfonic acid (MOPS) (free acid)

50 mM sodiumacetate

10 mM EDTA

pH to 7.0 withNaOH

1x FormaldehydeAgarose Gel Running Buffer

100 ml 10x FormaldehydeAgarose gel buffer

20 ml 37%(12.3 M) formaldehyde

880 mlRNase-free water

5x RNALoading Buffer

16 µlsaturated aqueous bromophenol blue solution†

80 µl 500 mMEDTA, pH 8.0

720 µl 37%(12.3 M) formaldehyde

2 ml 100%glycerol

3084 µlformamide

4 ml 10 x FormaldehydeAgarose gel buffer

RNase-freewater to 10 ml

Stability:Approximately 3 months at 4°C

Ref

2M H2SO4 for 100ml

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2M H₂SO₄ for 100ml

mix 50ml Milli Q water + 50ml 4M H₂SO₄ solution

4M H2SO4 for 1 liter

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4M H₂SO₄ for 1 liter

Carefully pour 392ml H₂SO₄ {Sulphuric acid 96% H₂SO₄ Carlo Erba reagents Code no. 410301 (Reanal 17769) FW 98.078} into 608ml Milli Q water (always pour acide into water)

TMB-buffer for 1 liter

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TMB-buffer for 1 liter[0.1M Na-acetate, pH 5.5]

Disolve in 950ml Milli Q water

13.6g Na-acetate {Nátrium acetát, kristályvizes CH₃COONa*3H₂O M:136.08 Reanal 14021}

Adjust pH to 5.5 with app. 700ul cc. Acetic acid {Ecetsav 96% Reanal 0914-1-08-65 M:60.05}
Adjust volume to 1 liter with Milli Q water

TMB solution for 1ml - ELISA

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TMB solution for 1ml [10mg/ml TNB in DMSO]

Disolve in 1ml DMSO {Dimethyl sulfoxide, minimum 99.5% GC Sigma D5879-100ml }

10mg TNP {3,3',5,5'-tetramethyl-benzidine Sigma T-2885 FW240.3}

Store at +4C

ELISA blocking-buffer for 10ml

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ELISA blocking-buffer for 10ml [0.05% Tween 20, 5%BSA, 0.05% azide, PBS]

Mix on magnetic stirrer:

10ml PBS-Tween
100mg BSA {Sigma, No.:A3059-100G, Albumin, bovine serum, Fraction V}
50ul Sodium azide (from 10% NaN₃ stock solution)

Store at +4C

Carbonate buffer pH 9.5 for 1 liter (for ELISA coat)

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Carbonate buffer pH 9.5 for 1 liter (for ELISA coat) Disolve in 1 liter Milli Q water: 1.6g Na2CO3 2.9g NaHCO3 Adjust to pH 9.5 Store at R/T

ELISA Method

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Method Coat ELISA plate with 50ul/well 0.2-10ug/ml protein (purity should be above 3%) in PBS or carbonate buffer for 2h on R/T or O/N in the fridge Wash 3x with PBS-Tween (alternative: following washing incubate in ELISA-blocking buffer for 30min R/T and wash 3x with PBS-Tween) Incubate in PBS-Tween diluted antibody/protein solution for 1h at 37C (ideally after 5h reach the equibrium) Wash 3x with PBS-Tween Develope with TMB Add 100ul TMB developing solution to wells For 1 ELISA plate mix: 11ml TMB buffer 110ul TMB solution 22ul H2O2 {Hydrogen peroxide 30% solution Sigma H-1009} Add 100ul 2M H2SO4 to stop the reaction Detect absorbance at 450nm

5x Veronal buffer for 500ml (5xVBS) Recipe

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5x Veronal buffer for 500ml (5xVBS) [727mM NaCl, 9.12mM Na-diethyl-barbiturate (C8H11O3N2Na), 15.63mM 5,5’-diethyl barbiture acid (C8H12N2O3), pH7.3] Make solution 'A': 0.94g Na-diethyl-barbiturate (C8H11O3N2Na){5,5-Diathylbarbitursaure. Na-salz Serva 18797; MW:206.2} 21.25 g NaCl {Reanal, No.:2464-1-22-38, Nátrium-klorid, Ph. Eur.5; MW:58.44} ~300ml Milli Q water Make solution 'B': 1.44g 5,5’- diethyl barbiture acid {5,5-dietil-barbitursav Reanal 04049; MW:184.2} ~100ml Milli Q water Boil it otherwise it won’t dissolve, then cool it down Mix solution ‘A’ and ‘B’ set pH to 7.3 with app. 50-70μl 10N NaOH Fill up until 500ml with Milli Q water, then store it at +4C

1x Veronal buffer recipe

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1x Veronal buffer (VBS)[145mM NaCl, 1,8mM Na-diethyl-barbiturate (C8H11O3N2Na), 3mM 5,5’-diethyl barbiture acid (C8H12N2O3), pH7.3] Dilute 5x Veronal buffer with Milli Q water

0.2M EGTA stock solution for 10ml

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0.2M EGTA stock solution for 10ml Mix on magnetic stirrer: 8ml Milli Q water 0.7608g EGTA{ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) Sigma E-4378 LOT 111K5411 FW 380,4} Adjust to pH8 with 10N NaOH otherwise it won't disolve (be careful EGTA is an acid) Adjust volume to 10ml with Milli Q water Store at 4C

EDTA-VBS solution for 10ml

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EDTA-VBS solution for 10ml (dilute serum 5x with it)[25mM EDTA, 5%BSA, 0.05% Tween 20, VBS ] Mix on magnetic stirrer: 2ml 5x VBS 500ul 0.5M EDTA 0.5g BSA {Sigma, No.:A3059-100G, Albumin, bovine serum, Fraction V} 50ul 10% Tween 20 {Sigma No.:P-1379, Polyoxylethylene-sorbitan monolaurate (Tween 20)} Adjust volume to 10ml with Milli Q water Filtrate though an 0.45um filter Aliquot 1ml/eppendorf Store at -20C

Mg-EGTA-VBS solution for 10ml

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Mg-EGTA-VBS solution for 10ml (dilute serum 5x with it)[2.5mM Mg2+, 6.2mM EGTA, 5%BSA, 0.05% Tween 20, VBS] Mix on magnetic stirrer: 2ml 5x VBS 310ul 0.2M EGTA 25ul 1M MgCl2 stock solution 0.5g BSA {Sigma, No.:A3059-100G, Albumin, bovine serum, Fraction V} 50ul 10% Tween 20 {Sigma No.:P-1379, Polyoxylethylene-sorbitan monolaurate (Tween 20)} Adjust volume to 10ml with Milli Q water Filtrate though an 0.45um filter Aliquot 1ml/eppendorf Store at -20C

Ca-Mg-VBS (veronal buffered saline) for 25ml

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Ca-Mg-VBS (veronal buffered saline) for 25ml (for serum dilution)[2.5mM Ca2+, 0.7mM Mg2+, 0.05% Tween 20, 5%BSA, VBS] Mix on magnetic stirrer: 5ml 5x VBS 250ul Ca2+ Mg2+ stock solution [0.25M Ca2+, 0.07M Mg2+] 1.25g BSA {Sigma, No.:A3059-100G, Albumin, bovine serum, Fraction V} 12.5ul Tween 20 {Sigma No.:P-1379, Polyoxylethylene-sorbitan monolaurate (Tween 20)} Adjust volume to 25ml with Milli Q water Filtrate though an 0.45um filter Aliquot 1ml/eppendorf Store at -20C

Ca2+ Mg2+ stock solution for 5ml [0.25M Ca2+, 0.07M Mg2+]

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Ca2+ Mg2+ stock solution for 5ml [0.25M Ca2+, 0.07M Mg2+] Dissolve in 5 ml Milli Q water: 0.1838g CaCl2*2H2O {Reanal 11024 MSZ 24161-65 Kalcium-klorid, szárított MW: 147.02 CaCl2*2H2O} 0.0712g MgCl2*6H2O {Reanal No.: 20281-1-01-38 Magnézium-klorid 6-hidrát MW: 203.30 MgCl2*6H2O}

PBS-Tween BSA for 300ml [0.05% Tween 20, 5%BSA, 0.05% azide, PBS]

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PBS-Tween BSA for 300ml [0.05% Tween 20, 5%BSA, 0.05% azide, PBS] Mix on magnetic stirrer: 300ml PBS-Tween 15g BSA {Sigma, No.:A3059-100G, Albumin, bovine serum, Fraction V} 1.5ml Sodium azide (from 10% NaN3 stock solution) Filtrate through an 0.45um filter Aliquot 50ml/falcon Store at 4°C

PBS-Tween for 1 liter [0.05% Tween 20, PBS]

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PBS-Tween for 1 liter [0.05% Tween 20, PBS] Mix on magnetic stirrer: 0.5ml Tween 20 {Sigma No.:P-1379, Polyoxylethylene-sorbitan monolaurate (Tween 20)} 1 liter PBS

1x PBS Recipe for 1 liter[137mM NaCl, 2.7mM KCl, 8mM Na2HPO4, 1.46mM KH2PO4]

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Dissolve in 800 ml distilled water: 8 g NaCl {Reanal, No.:2464-1-22-38, Nátrium-klorid, Ph. Eur.5; MW:58.44} 0.2 g KCl {Reanal, No.:18050-1-01-38, Kálium-klorid, a.r.; MW: 74.56} 1.424 g Na2HPO4*2H2O {Reanal, No.:08973-1-01-38, di-Nátrium-hidrogén-foszfát 2-hidrát, a.r.; MW:177,99} 0.2 g KH2PO4 {Reanal, No.:17890-01-38, Kálium-dihidrogén-foszfát, a.r.; MW:136.09} Adjust volume to 1 liter with H2O Filtrate trough an 0.22um filter or sterilize by autoclave Store at room temperature

10x PBS Recipe for 1 liter

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10x PBS Recipe for 1 liter Dissolve in 800 ml distilled water: 80 g NaCl {Reanal, No.:2464-1-22-38, Nátrium-klorid, Ph. Eur.5; MW:58.44} 2 g KCl {Reanal, No.:18050-1-01-38, Kálium-klorid, a.r.; MW: 74.56} 14.24 g Na2HPO4*2H2O {Reanal, No.:08973-1-01-38, di-Nátrium-hidrogén-foszfát 2-hidrát, a.r.; MW:177,99} 2 g KH2PO4 {Reanal, No.:17890-01-38, Kálium-dihidrogén-foszfát, a.r.; MW:136.09} Adjust volume to 1 liter with H2O Filtrate through an 0.22um filter and divide in 500ml aliquotes Store at room temperature

RNA Isolation Protocol from Tissue Samples

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Place the desired tissue in the prechilled mortar, add liquid nitrogen and grind to a fine powder with the pestle... Transfer the ground tissue to a Falcon 2059 tube and add 1ml monophasic isolation reagent per 100mg tissue... Homogenize the tissue for 15-30 seconds with a polytron mechanical homogenizer... Incubate the homogenized sample for 5 minutes at RT to allow complete dissociation of protein complexes.Read Full protocol here.

Amyloid Beta HFIP Stock Protocol

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Lyophilized peptide should be ideally ordered in 10 mg glass vials. In-house material should beaccurately weighed in clean glass vials with a HFIP resistant closure.Read Full protocol Here.

Fluorimetric Enzyme Assay Protocol for Marine Samples

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http://allison.bio.uci.edu/Protocols/EnzymeProtocolMarine.pdf

GRAM STAIN PROTOCOL - fecal smear to observe the bacterial microflora

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This technique is used to stain a slide such as a fecal smear to observe the bacterial microflora present based on their gram stain reaction.“Heat-fix” the slide with the specimen by passing it over a heat source, such as aflame, several times using a forceps. The slide should be passed very quickly throughthe flame and not be heated excessively. Place slide on the staining tray.Read Full Protocol Here

HIV/AIDS Vaccine Developed in Canada

2011-12-20T17:07:00.001-08:00

HIV/AIDS vaccine developed at Western proceeding to human clinical trials. After two decades of research, a group of Canadian scientists has won approval to start testing an experimental HIV vaccine on humans.

How Bubble Gum is Made?

2011-12-16T10:33:00.000-08:00

Check this out. If you don't know how Gum is made.

How to make 1x Trypsin

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Use the formula C1V1=C2V2 to make the dilution for your required volume.

C=Concentration
V=Volum

EXAMPLE
Make 20 ml of 1X Trypsin from a 10X stock.

        10X stock *  X  =  1X stock *  20 ml

                     X = (1X stock *  20 ml) / 10X stock

                     X = 2.0 ul 10X stock
                       +18.0 ul of PBS

How to make 1x TAE Buffer

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To make 1X TAE take 20mL of 50X TAE and fill up the remaining volume of a
1000mL erlenmeyer flask with nano-pure water.

How to make 1x tae from 50x tae

Add 1ml of 50x to 49 ml of water. Increase the volume as required.

How to make 1x pbs from 10x pbs

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If you have tablets then make the 10x pbs solution according the the manufacturer's instructions.

Than use the formula C1V1=C2V2 to make the dilution for your required volume.
C=Concentration
V=Volum

Make 20 ml of 1X PBS buffer from a 10X stock.

        10X stock *  X  =  1X stock *  20 ml

                     X = (1X stock *  20 ml) / 10X stock

                     X = 2.0 ul 10X stock
                       +18.0 ul of H₂O

how to make 1x from 50x

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You need to use the formula

C1V1=C2V2
C=Concentration
V=Volume

Example

Make 20 ul of 1X restriction enzyme buffer from a 10X stock.

        10X stock *  X  =  1X stock *  20 ul

                     X = (1X stock *  20 ul) / 10X stock

                     X = 2.0 ul 10X stock
                       +18.0 ul of H₂O

How to make 1x from 10x

2011-12-13T16:38:00.000-08:00

Simple Way to make Easy Dilutions - how to make 1x solution from 10x solution

10x means 10 Times concentrated. If you want to make 1x then you have to dilute it 10 time. For Example. To make 10 ml of 1x solution in water.You need take 1 ml of solution [10px] and mix with 9 ml of water. Final total volume is now 10 ml [10 times of 1ml]Done :)

Theoretical Way to make dilutions

You should use the formula C_i V_i = C_f V_f
_or

_M1V1=M2V2

Example

Make 20 ul of 1X restriction enzyme buffer from a 10X stock.

        10X stock *  X  =  1X stock *  20 ul

                     X = (1X stock *  20 ul) / 10X stock

                     X = 2.0 ul 10X stock
                       +18.0 ul of H₂O

Amit

Isolation of Bacteria from the Environment - References

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Isolation of bacteria from the environmenthttp://delrio.dcccd.edu/jreynolds/microbiology/2420/files/bacteria_ubiquity.pdf

Laboratory Notes for BIO 1003 John H. Wahlert & Mary Jean HollandExamination of dental bacteriahttp://faculty.baruch.cuny.edu/jwahlert/bio1003/eubacteria.html

Culture and isolation of phototrophic sulfur bacteria from the marine environmenthttp://adsabs.harvard.edu/abs/1970HWM....20....6T

Isolation of Bacteria from Soil Samples

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ISOLATION OF SOIL BACTERIA: VIABLE TITER and PURE CULTURE
http://www2.fiu.edu/~makemson/MCB2000Lab/Exp4SoilBact.pdf

Isolation of an Unknown Bacterium from Soil
Department of Biological Sciences
University of Nevada, Las Vegas
http://www.ableweb.org/volumes/vol-14/6-steubing.pdf

ISOLATION OF PHOSPHATE SOLUBILISING BACTERIA FROM SOIL AND ITS ACTIVITY
http://biotechindia.files.wordpress.com/2007/12/isolation.pdf

Isolation of Soil Microorganisms
http://apps.caes.uga.edu/sbof/main/lessonPlan/microorganismIsolation.pdf

Isolation of bacteria from mechanic workshops’ soil environment contaminated with used engine oil
Department of Medical Laboratory Sciences, College of Medicine, University of Nigeria, Enugu Campus, Enugu, Nigeria.

Isolation of soil Streptomyces as source antibiotics active against antibiotic-resistant bacteria
http://ejobios.com/pdf/ejob-8-11-2,9,73-82.pdf

ISOLATION OF SOIL BACTERIA FOR BIOREMEDIATION OF HYDROCARBON CONTAMINATION
http://www.chem.msu.su/rus/vmgu/031/88.pdf

RNA isolation from soil for bacterial community and functional analysis: evaluation of different extraction and soil conservation protocols
http://www.engr.colostate.edu/~apruden/classes/ce742/Readings/Articles/RNAextractTechniques.pdf

TEM-PCR Method,Technology and Testing

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What is TEM-PCR Technology?

The TEM-PCR stands for Target Enriched Multiplexing Polymerase Chain Reaction. TEM-PCR is a molecular multiplex PCR technology used Molecular Differential Detection. By TEM-PCR the molecular targets can be amplified in one reaction. This technology is advantageous over the RT-PCR Technology because multiple molecular targets can be amplified specifically in a single reaction. This makes it a important tool for detection of molecular markers of pathogens.

In last few years improvements have been done in this technology. Benson et. al. compared the ResPlex I assay kit of Qiagen with RT-PCR in the detection of various bacterial strains of respiratory disease and found that it is less sensitive than real-time PCR but has advantage over multiple assays [ref]. Deng et. al. detected more S. pneumoniae (32 vs. 7) and H. influenzae (29 vs. 23) by TEM-PCR than did culture [ref].Related:Overlap Extension PCR

Isolation of Actinomycetes

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References for Isolation of Actinomycetes

Culturable marine actinomycete diversity from tropical Pacific Ocean sediments.

Jensen PR, Gontang E, Mafnas C, Mincer TJ, Fenical W.

Source

Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California - San Diego, La Jolla, CA 92093-0204, USA
http://www.ncbi.nlm.nih.gov/pubmed/15946301

Isolation and characterization of a sponge-associated actinomycete that produces manzamines

Marine-Estuarine-Environmental Sciences

http://drum.lib.umd.edu/handle/1903/4114

Isolation of Acid Phosphatase

2011-12-11T16:08:00.001-08:00

Here we have searched and compiled a list of methods used by various college and universities for the isolation of acid phosphatase.

Purification of Acid Phosphatase from Wheat Germ
There are three major steps involved in the purification of macromolecules. First, you must disrupt the cell you are using as a source. Second, you must selectively purify the macromolecule away from contaminating molecules. Third, you must both prevent degradation of the macromolecule during the procedure and you must preserve the native structure of the macromolecule to be purified. In the purification of proteins, you want to separate that protein from other “contaminating” proteins, and you want to purify it in its native conformation with retention of enzymatic activity.
Further Details

Purification and Partial Characterization of an Acid Phosphatase fromSpirodela oligorrhiza and Its Affinity for Selected Organophosphate Pesticides

Christopher F. Hoehamer,^†^‡ Chris S. Mazur,*^‡ and N. L. Wolfe^‡

National Research Council, and National Exposure Research Laboratory, U.S. Environmental Protection Agency, 960 College Station Road, Athens, Georgia 30605http://pubs.acs.org/doi/abs/10.1021/jf040329u

Isolation and partial characterization of an acid phosphatase

from Artemisia vulgaris pollen extract

TANJA D. ]IRKOVI]*#, MARIJA DJ. GAVROVI]-JANKULOVI]* #, MIRJANA N. BUKILICA**,

LJUBA MANDI]*#, SPOMENKA Z. PETROVI]*** and RATKO M. JANKOV*

http://www.doiserbia.nb.rs/img/doi/0352-5139/2002/0352-51390209567C.pdf

Isolation of Acetylsalicylic Acid from Aspirin Tablets

2011-12-11T16:01:00.001-08:00

Here we have compiled a list of laboratory protocols from various colleges and universities for the Isolation of acetylsalicylic acid from aspirin tablets.

Isolation of acetylsalicylic acid from aspirin tablets

Organic Chemistry Laboratory CHE 327

Cornell College Term 7, 2004/2005

Addison Ault, Andrea Pionek, Mary Anne Teague, Charley Liberko

http://people.cornellcollege.edu/cliberko/OrgLabManual.htm

Organic Chemistry Laboratory
Isolation of Acetysalicylic Acid from Aspirin Tablets

Aspirin, like most tablets, contains the active ingredient (acetylsalicylic acid) plus a binder to give the tablet its shape. The active ingredient in aspirin will be recovered by recrystallization from ethanol. The aspirin tablets you will be using contain 325 mg of acetylsalicylic acid per tablet. (However, check the bottle to be certain, as occasionally substitutions have to be made.)

From Mansfield.edu [PDF]

Synthesis of Acetylsalicylic Acid (Aspirin)

Introduction: The purpose of this experiment is to synthesize acetylsalicylic acid via an esterification
reaction between salicylic acid and acetic anhydride. The product will be recrystallized using
95% ethanol. The product will then be analyzed via infrared spectroscopy, nuclear magnetic
resonance spectroscopy, ultraviolet light visible spectroscopy, and melting point analysis. It
will then be compared with the IR, NMR, and UV spectra and melting point of acetylsalicylic
acid isolated from commercial aspirin
http://www.franklincollege.edu/pwp/lmonroe/Organic%20Chem/Aspirin.pdf

Lysis buffer: RIPA or NP40 for Immunoprecipitation

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Lysis buffer.

The choice of lysis buffer depends on what kind of IP you are doing.

RIPA buffer gives the lowest background, but can denature some kinases. It also has the potential to disrupt protein:protein interactions.

NP40 buffer (EVF) is less denaturing, but gives a higher background. It is less likely to inhibit kinase activity and disrupt protein complexes.

RIPA Buffer recipe

Further Buffer Information

RIPA Lysis Buffer Recipe : RIPA buffer for mammalian cell lysis:

2011-12-11T15:50:00.001-08:00

RIPA buffer for mammalian cell lysis:
50mM Tris-cl pH 7.4
150mM NaCl
1% NP40
0.25% Na-deoxycholate
1mM PMSF
1x Roche complete mini protease inhibitor cocktail
1x Pierce phosphatase inhibitor cocktail

Stock solutions used:
100mM Tris-Cl pH 7.4, 300mM NaCl
10% NP40 in water
10% Na-deoxycholate in water (stored at RT in dark)
200mM PMSF in isopropanol
Roche complete protease inhibitor cocktail tablets (cat# 04 693 124 001)
Pierce phosphatase inhibitor cocktail (100x) (cat # 78420)
For 10ml of lysis buffer:
5ml 100mM Tris-Cl pH 7.4, 300mM NaCl
1ml 10% NP40 in water
3.65ml water
250ul 10% Na-deoxycholate in water
1 tablet * Roche complete protease inhibitor cocktail
100ul* Pierce phosphatase inhibitor cocktail
50ul* 200mM PMSF in isopropanol

*add at time of use. PMSF is only good for ~30minutes after dilution into buffer.

Reference [pdf]

RIPA Buffer Recipe

2011-12-09T13:28:00.001-08:00

EDTA-free RIPA (RIPA[-]):
0.1% NP40, 20mM Tris/HCL pH
8,10% Glycerol, store at 4ºC (Usually
RIPA buffer contains 1 mM EDTA

Reference

10x PBS Buffer Recipe

2011-12-09T13:25:00.001-08:00

10X PBS BUFFER
To make 1 Liter

1. Dissolve the following in ~800mL18 MΩ dH2O. May need to heat a little to dissolve completely.
80g NaCl 14.4g Na2HPO4–7H2O (dibasic)
2g KCl 2.4g KH2PO4 (monobasic)
2. Adjust pH to 7.4 with HCl or NaOH.
3. Raise volume to 1 liter.
4. Transfer to a 1L bottle, label and autoclave.

Reference

10X, 20X TRIS Buffer Recipes

2011-12-09T13:22:00.001-08:00

10X Tris-HCl (0.5M Tris Base, pH7.6):
Trizma Base ---------------------------------- 61 g
Distilled water ------------------------------- 1000 ml
Adjust pH 7.6 using concentrated HCl
Store this solution at room temperature. Dilute 1:10 with distilled water before use
and adjust pH if necessary.

20X Tris-HCl (1M Tris Base, pH7.6):
Trizma Base ---------------------------------- 122 g
Distilled water ------------------------------- 1000 ml
Adjust pH 7.6 using concentrated HCl
Store this solution at room temperature. Dilute 1:20 with distilled water before use
and adjust pH if necessary.

10X Tris-HCl-Tween 20 (0.5M Tris Base, 0.5% Tween 20, pH7.6):
Trizma Base ---------------------------------- 61 g
Distilled water ------------------------------- 1000 ml
Adjust pH 7.6 using concentrated HCl and then add 5 ml of Tween 20.
Store this solution at room temperature. Dilute 1:10 with distilled water before use
and adjust pH if necessary.

Reference

10% SDS Recipe

2011-12-09T13:15:00.001-08:00

10% SDS Recipe

1L:

100g SDS into 1 L, heat to 68C for solubility. pH ~6.6.

50X TAE DNA Electrophoresis Buffer Recipe

2011-12-09T13:14:00.001-08:00

TAE DNA Electrophoresis Buffer (50 X)

(2 M Tris, 50 mM EDTA)

2 L
484 g Tris
114.2 ml glacial acetic acid
200 ml 0.5 M EDTA 8.0
To make 1x TAE 20 L, add 400 ml 50X buffer into 19.6 L ddH2O

Reference

0.5M EDTA pH 8.0 Recipe

2011-12-09T13:13:00.001-08:00

EDTA 0.5 M (pH8.0)

0.5M, 1L: 148 g EDTA
+ ~30-40 g NaOH to adjust pH
(or 186 g EDTA-Na.2H2O + ~20 g NaOH)
Note: pH adjusted by NaOH is essential for solubility. Autoclavable

Reference

1 M TRIS Buffer Recipe

2011-12-09T13:11:00.001-08:00

1 M Tris-HCl Buffers

pH Volume (L) TrisBase (g) HCl (ml)
pH 7.0 2 242.2 150-155
pH 7.5 2 242.2 120-125
pH 8.0 2 242.2 80-85

Autoclavable

Reference

0.5 x TBE Buffer Recipe

2011-12-09T13:04:00.001-08:00

0.5 x tbe buffer recipe

-108 g Tris base
-55 g Boric acid
-40 ml 0.5 M EDTA pH 8.0
-H2O to 2 L

Place the flask on a magnetic stirrer until completely dissolved. This results in a 5X concentrated
stock of TBE Buffer. Dilute the 2L of 5X stock by 10 in distilled water (add 18 L or 4.5 gallons of
distilled water). Mix well. The buffer is ready for use.

Reference

Career Fair at EuroBio 7-9 October in Paris

2008-07-08T03:06:00.001-07:00

EuroBiO Career Fair is a three day event 7-9 October in Paris, that benefits from the presence of more than five thousand participants in EuroBiO’s other Pillars (121 business partnering, a lobbying platform and an exhibition) to generate optimum market conditions for recruiters and job seekers.
It’s aimed at recruiters, job seekers and higher education establishments and seeks to further the mobility of researchers throughout the European Research Area.

This year the Career Fair has partnered with the London Biotechnology Network, Association Bernard Gregory and ScienceCareers to put on a special Career Fair Open Day to be held on Wednesday 8th October at BioCity, a dedicated careers area inside the EuroBiO Exhibition. The Career Fair Open Day will put the accent on alternative careers (to the laboratory), and the multiple talents that biotechnology companies require to grow their business. Candidates with the minimum of a Masters Degree in Sciences (or final year PhD) will be able to introduce themselves to recruiters, without prior appointment, and recruiters subsequently will be able to offer one-to-one interviews in booths to those profiles that correspond to their requirements. The Open Day seminars will be looking at alternative science careers such as journalism, degrees for managing biotechnology companies, and concrete measures that facilitate the mobility of researchers. Candidate registration also includes access to the exhibition which showcases Life Sciences stakeholders and leading BioClusters from across the world, the challenging EuroBiO Plenaries, the forward-looking Presenting Company sessions... and networking breaks!

Don't stop your microarray research this summer

2008-07-02T09:16:00.001-07:00

Don’t stop your microarray work this summer.

Use a fluorescent dye that has:

	Reproducible performance all year round
	3-fold more ozone stability
	0% more photostability

Amersham HyPer5 dye is a new fluorescent dye for microarray applications. It combines exceptional ozone and photostability with highly reproducible performance all year round—irrespective of environmental conditions.

To learn more about the exceptional performance of Amersham HyPer5 download a copy of our "Evaluation on the performance of Amersham HyPer5™ dye in comparative genomic hybridization microarray applications" from the Center of Human Genetics, Belgium.

Take a moment and register to download your free copy.

http://www.promo.gelifesciences.com/na/k8038nawx/reg.asp

New Snap Tube E-Blast

2008-06-27T02:40:00.000-07:00

Finally a way to make your PCR or QPCR secure and simple. The Thermo Scientific ABgene EasyStrip Snap Tubes are 8-well strip tubes with individually sealable caps. Each individual cap will eliminate any chance of cross contamination and make your workflow much easier and more secure.

Optimized Capping Options – Application specific strips available with ultra clear caps for QPCR and domed caps for PCR.

Increased Sensitivity in QPCR – white tubes and Ultra Clear Caps have been proven to increase detection of low copy number templates.

Eliminate Cross Contamination – Individually attached caps do not create aerosols, unlike strip caps, when they are removed after cycling.

EasyStrip Snap Tube sample request page

http://www.abgene.com/snapsample.asp

Advanced Course in Immunology University of Minnesota

2008-06-19T09:01:00.001-07:00

Advanced Course in Immunology

July 19-24, 2008
University of Minnesota/Minneapolis Campus
Course Directors: Marc K. Jenkins, Ph.D. and
Kristin A. Hogquist, Ph.D., University of Minnesota
Center for Immunology

REGISTRATION OPEN
http://www.aai.org/Adv_Course/2008/Program.htm
Deadline: Monday, June 30, 2008!

This intensive one-week course is designed for serious students of
immunology. Leading experts will present recent advances in
understanding the biology of the immune system and its role in health
and disease.

The course is intended for scientists and advanced trainees who wish to
expand or update their understanding of the field. This is not an
introductory course and requires that attendees have a firm
understanding of the principles of immunology.

Accommodations at special course rates have been arranged at the
Radisson University Hotel.
http://www.radisson.com/aai2008

Applicants from outside the US are encouraged to apply early for visas.

The AAI Courses in Immunology are sponsored by The American Association
of Immunologists, the world's largest and most prestigious association
of professional scientists engaged in basic and clinical immunological
research.

University of Minnesota/Minneapolis Campus, site of the AAI 2008
Advanced Course in Immunology

http://www.aai.org/Adv_Course/2008/2008AdvCourse.04.28.08HighRes.pdf

Microporation: Low cost of electroporation

2008-06-19T06:56:00.001-07:00

User friendly interface

High transfection efficiency and good cell survival rate

Plasmid, protein and SiRNA transfection

Low cost of electroporation

Microporation is a novel and proprietary electroporation technology developed by Digital Bio Technology. Before the Microporation, all of the electroporation is dependent on the plate-type electrode and cuvette style chamber. Microporation is a unique electroporation technology using a pipette TIP as an electroporation space. In doing so, the uniform electric field is generated accompanying minimal heat generation, metal ion dissolution, pH variation and oxide formation ‥‥‥more <http://www.microporator.com/introduction/intorduction_1.php>

MegaTran 1.0: the transfection reagent for large volume applications

2008-06-16T03:24:00.001-07:00

MegaTran 1.0: the transfection reagent for large volume applications -
10 ml for $1200

MegaTran 1.0 is a brand new non-lipid polymer transfection reagent
specially designed and manufactured for large volume DNA transfection,
including large scale protein production via transient transfection,
high-throughput screening using cDNA arrays or shRNA libraries, etc.

Superior transfection efficiency compared to leading transfection
reagents
Improved protein production via transient transfection
Reduced cytotoxicity
Extremely affordable - $1200 for 10 ml!

Further Information Here
http://www.origene.com/cdna/megatran1.mspx

Reminder: Webinar Invitation - Advances in GPCR Research, June 17

2008-06-05T07:29:00.001-07:00

Advances in GPCR Research WEBINAR

In case you missed the first announcement, or didn’t have the opportunity to register yet, we wanted to remind you to please join our panel of experts on 17 June 2008 for a live, online educational seminar, "Advances in G Protein-Coupled Receptor Research."

Submit your questions LIVE to the experts during the webinar!

Date: Tuesday, June 17, 2008
Time: 12:00 noon Eastern Time; 9:00 a.m. Pacific Time; 4:00 p.m. GMT
Duration: 1 hour

Register now! For more information and complimentary registration visit:
www.sciencemag.org/webinar

Membrane bound G protein-coupled receptors (GPCRs) play an integral role in sensing the external environment of the cell. A broad range of external stimuli signal through GPCRs, including neurotransmitters, hormones, odor molecules, cations, and even photons. Their position at the apex of essential signal transduction pathways means that malfunctioning of these molecules frequently leads to disease, making them a perfect target for drug therapies. Further, naturally occurring and induced mutations in GPCRs can provide valuable information about these signaling pathways and their role in human pathologies.

You are invited to join our panel of thought leaders in a live video webinar discussion about these very important proteins. During the broadcast, you will:

*       gain insight into the broad range of GPCR proteins and their role in signaling and disease
*       learn about techniques and technologies available to study GPCRs
*       see data from recent GCPR studies, presented by top scientists in the field
*       have the opportunity to ask questions of the expert panel LIVE during the event

Participants:

Jeffrey Conn, Ph.D.
Vanderbilt University Medical Center
Nashville, TN

Michel Bouvier, Ph.D.
Institute for Research in Immunology and Cancer (IRIC)
Montreal, Canada

Brian Kobilka, M.D.
Stanford University School of Medicine
Stanford, CA

REGISTER NOW AT: www.sciencemag.org/webinar

Advances in GPCR Research WEBINAR

2008-05-21T09:09:00.003-07:00

Advances in GPCR Research WEBINAR

Tuesday, June 17, 2008, for a live, online educational seminar entitled, "Advances in G Protein-Coupled Receptor Research."

Submit your questions LIVE to the experts during the webinar!

Date: Tuesday, June 17, 2008
Time: 12:00 noon Eastern Time; 9:00 a.m. Pacific Time; 4:00 p.m. GMT
Duration: 1 hour

You are invited to join our panel of thought leaders in a live video webinar discussion about these very important proteins. During the broadcast, you will:
• gain insight into the broad range of GPCR proteins and their role in signaling and disease
• learn about techniques and technologies available to study GPCRs
• see data from recent GCPR studies, presented by top scientists in the field
• have the opportunity to ask questions of the expert panel LIVE during the event

Participants:

Jeffrey Conn, Ph.D.
Vanderbilt University Medical Center
Nashville, TN

Michel Bouvier, Ph.D.
Institute for Research in Immunology and Cancer (IRIC)
Montreal, Canada

Brian Kobilka, M.D.
Stanford University School of Medicine
Stanford, CA

Eppendorf epMotion Plug'n'Prep - Automate ANY Nucleic Acid Purification Protocol in Seconds

2008-05-21T09:09:00.001-07:00

Because epMotion is an open system, you are free to choose the type of extraction kits you need from the manufacturers that you prefer.

And since epMotion can fully automate your protocol, you are also free to do other tasks while your sample is being purified. All epMotion methods have been optimized and tested in cooperation with the kit manufacturer. Our Plug'n'Prep protocols provide superior nucleic acid yield and purity.

For more information visit www.epMotion.com/pnp

Dig deeper in the proteome with ProteoMiner kits!

2008-04-11T02:27:00.001-07:00

Dig deeper in the proteome with ProteoMiner kits!

ProteoMiner protein enrichment kits enrich and concentrate low-abundance
proteins that cannot
be detected through traditional methods. The kits include spin columns
and all necessary reagents for detecting low-abundance proteins,
utilizing
a simple, easy-to-use procedure, which allows for parallel processing of
samples. The kits can be used with a variety of biological samples and
are
compatible with all major downstream proteomics applications. For more
information, visit

www.bio-rad.com/proteominer/

New DUALmembrane kit

2008-03-28T03:45:00.001-07:00

Find novel interaction partners for your integral membrane protein!
With the DUALmembrane kit you can screen full-length, integral membrane proteins and membrane-associated proteins against a cDNA library of your choice to identify novel interaction partners of your protein of interest.

Features of the DUALmembrane kit 3
Screen full-length integral membrane proteins for interactors in vivo
Protein interactions are detected under physiological conditions at the membrane
Novel interactors are identified through screening of cDNA libraries
Easy subcloning of full-length cDNA inserts using Sfi I technology
NMY51 reporter strain with improved stringency, resulting in fewer false positives
Trial size HTX β-galactosidase assay kit included (2x 96 reactions)

DUALmembrane starter package
Take advantage of the DUALmembrane starter package to get quickly started: it includes the DUALmembrane kit 3 and two additional kits for easy quantitative determination of β-galactosidase levels and efficient library scale yeast transformation.

The DUALmembrane starter package includes:

DUALmembrane kit 3
Standard size HTX β-galactosidase assay kit (4x 96 reactions)
DS Yeast transformation kit

Further information
http://www.dualsystems.com/index.php?id=86

New qRT-PCR Solutions from Applied Biosystems

2008-03-22T11:36:00.000-07:00

qRT-PCR Solutions from Applied Biosystems!
We are pleased to announce our new suite of products that streamline reverse transcription reactions with improved performance, reliability and efficiency. All Applied Biosystems kits are optimized with our industry-leading TaqMan® Gene Expression Assays or our high-performance SYBR® Green reagents.

Move your reverse transcription technologies forward today!

High Capacity RNA-to-cDNA Master Mix for high throughput
High Capacity RNA-to-cDNA Kit for standard throughput
TaqMan® RNA-to-CT™ 1-Step Kit (Coming soon)
Power SYBR® Green RNA-to-CT™ 1-Step Kit
TaqMan® RNA-to-CT™ 2-Step Kit
Power SYBR® Green RNA-to-CT™ 2-Step Kit

For introductory offer and to select the kit that's right for you, try our qRT-PCR Decision Tree.

For more information, visit www.appliedbiosystems.com/qrtpcr

Bioline Competent Cells - Buy 1 Get 1

2008-03-11T07:15:00.001-07:00

Bioline Competent Cells

Alpha-Select™ Bacteriophage T1-Resistant Gold Efficiency
Alpha-Select™ Bacteriophage T1-Resistant Silver Efficiency
Alpha-Select™ Bronze Efficiency
Alpha-Select™ Gold Efficiency
Alpha-Select™ Silver Efficiency
CH3-Blue 10⁸ Chemically Competent Cells
CH3-Blue 10⁹ Chemically Competent Cells
BIOBlue 10⁸ Chemically Competent Cells
BIOBlue 10⁹ Chemically Competent Cells
dam- dcm- Bacteriophage T1-Resistant
BL21
BL21 (DE3)
BL21 (DE3) pLysE
BL21 (DE3) pLysS
BL21 ComboPack

Order Now!

Tel: 888 257 5155 • Fax: 508 880 8993
Email: info@bioline.com • URL: www.bioline.com

$0.66 per bp for Synthetic Genes - Cloning via Email

2008-02-06T04:45:00.001-08:00

Cloning used to be straight benchwork, involving planning, repetitive
digestion, purification, ligation, transformation, screening, and
confirmation. This tedious process can now be bypassed by simply
emailing the sequence of your gene of interest to a precise and reliable
contractor, cloning by email to Gen Script!

http://www.biologyproject.net/gene_synthesis.html

Free up your valuable time and energy to invest in more creative tasks.
Send us the sequence of your gene and you will receive it in any vector
in as little as 12 business days.

Our advanced gene synthesis technology applies to all our gene
manipulation services:
1) Mutagenesis - for as little as $295 per mutation
2) Construction of mutant libraries
3) Construction of targeting vectors (knockout/knockin) - for as little
as $2,800 per construct

Gen Script Corporation - VWR Strategic Partner

Protein Characterization Utilizing Unique New Capabilities from a Dynamic Light Scattering Instrument

2007-12-11T17:28:00.001-08:00

Protein Characterization Utilizing Unique New Capabilities from a Dynamic Light Scattering Instrument
The molecular weight of biological materials and polymers can be determined from both static light scattering (SLS) and dynamic light scattering (DLS) experiments using Malvern's Zetasizer Nano. In addition the molecular size of very small molecules in solution can be measured using the Zetasizer Nano, both as a batch measurement in cuvettes, and with the use of a flow cell, as a detector at the end of a chromatography column.
more...

Cell Signaling: Chasing Down Kinases And Their Targets

2007-12-07T09:38:00.001-08:00

Cell Signaling: Chasing Down Kinases And Their Targets
Signaling pathways are everywhere. We couldn’t live without them – both the intracellular pathways that govern the interior workings of cells, and the pathways involved in intercellular communication, and the communication of cells with their physical environment. But these nicely organized... more...

No More Slab Gels for DNA and RNA

2007-12-07T09:37:00.001-08:00

No More Slab Gels for DNA and RNA
eGene Inc., now a Qiagen company, sells the HDA-GT12TM Genetic Analyzer for the separation and detection of DNA/RNA molecules. The HDA system automatically handles samples in 12-well strips or a 96-well tray, gives DNA resolution up to 2-5 bp, uses less than 1 µL sample volume, and provides DNA size and concentration data in digital format. This analyzer combines automatic gel electrophoresis and accurate gel documentation in one compact system.
more...

Get a head start on your writing this festive season

2007-12-05T03:02:00.001-08:00

Get a head start on your writing this festive season

If you're going to give the traditional Christmas telly a miss this year (how many times have you watched reruns of Only Fools and Horses or Mary Poppins!) to instead start work on your final dissertation or making serious headway on your thesis, you might want to think about treating yourself to a tool that will make creating citations and bibliographies a hundred times quicker and simpler. EndNote makes working with literature references easier at every stage, from capturing and organising relevant extracts, to using them in your writing.

Buy and download your copy of EndNote 24/7 (even on Christmas Day) to save the most money.

New EndNote case study - capturing and cataloguing palliative

2007-12-05T03:01:00.001-08:00

New EndNote case study - capturing and cataloguing palliative
care research

This case study shows how EndNote helped a team of researchers from the Association for Children's Palliative Care charity (ACT) gather together and organise relevant research to create a bi-annual journal called PaedPalLit. The journal delivers a complete, easy-to-reference guide to the latest palliative care research, providing an invaluable source of information to busy practitioners, clinicians and affected families.

Download your copy of the ACT EndNote case study

Free antibody production

2007-12-04T10:42:00.000-08:00

Visit www.novusbio.com and fill out the Antibody Grants Sheet for a chance to receive 2 mgs of antibody!

Grant Award Date: 1 Award selected on the 15th of every month. Awardees will receive a 0.2 mg test sample of affinity purified rabbit sera produced by Novus Biologicals. (Typical antibody production takes 4-5 months). If the product works and you supply the necessary documentation, you will receive 2 mgs of affinity purified antibody in exchange for product feedback.

Submit by the end of the month to be selected in the following month’s drawing.

Sign-up for future news on antibody grants,
new products and technical tips here

From Paraffin Sample to RT-ready RNA in 2 hours

2007-12-04T09:42:00.000-08:00

From Paraffin Sample to RT-ready RNA in 2 hours with Maximized Yield

WaxFree(tm) RNA is designed to extract RNA from both formalin-fixed
paraffin embedded (FFPE) samples and fresh and frozen tissue. The
straightforward protocol employs a non-binding method to efficiently
extract RNA. WaxFree minimizes the RNA loss typically observed in
traditional column or bead methods so it is particularly useful for old
or small samples. In addition, most extractions are completed in
approximately two hours without the use of toxic xylene.

Full details here
http://www.trimgen.com/product_rna.asp

PCRMatch.com Bio-Rad PCR and real-time PCR tools

2007-12-04T07:29:00.001-08:00

Bio-Rad is committed to providing the best tools for PCR and real-time PCR. Register now on PCRMatch.com and receive the information you'll need to find your perfect match.

Bio-Rad offers the most complete line of thermal cyclers available. With Bio-Rad cyclers
you can:

Quantitate up to 5 targets per well
Optimize annealing temperature in a single run
Increase success with enzymes that work where others fail
Improve reliability

Need more information? Visit the interactive PCR Doctor at www.bio-rad.com/genomics/support to define your PCR needs and learn the PCR tip of the week

Trypsin Recombinant, Proteomics Grade

2007-12-03T14:15:00.001-08:00

Trypsin Recombinant, Proteomics Grade
Roche's Trypsin Recombinant, Proteomics Grade is a highly purified protease especially designed for the digestion of proteins separated by 2-D gel electrophoresis or liquid chromatographic methods in the course of sample preparation for mass spectrometry. The enzyme is free of contamination from other proteases, and features high specific activity and a reproducible cleavage pattern, leading to faster digestions and more accurate identification of proteins in mass spectrometry. Roche's broad range of sequencing-grade proteases also includes Lys-C, Glu-C, Asp-N, Arg-C, Chymotrypsin, and Carboxypeptidase Y.
more...

Human Retinol-binding Protein 4 QuantikineR ELISA

2007-12-03T10:44:00.000-08:00

Human Retinol-binding Protein 4 Quantikine(r) ELISA
R&D Systems now offers a human Retinol-binding Protein 4 (RBP4)
Quantikine ELISA (Catalog # DRB400 ). RBP4 binds to and transports
retinol (vitamin A) from the liver to the peripheral tissues. RBP4 also
promotes hyperglycemia through downregulation of the glucose transporter
GLUT4 in adipocytes, the upregulation of the hepatic gluconeogenic
enzyme PEPCK, and the attenuation of insulin receptor signaling in
skeletal muscle. Serum RBP4 levels are also reported elevated in type 2
diabetes and obesity. This new ELISA is designed to measure RBP4
concentrations in cell culture supernatant, serum, plasma, urine, and
saliva. For more information, please visit www.rndsystems.com or call
1-800-343-7475.

Atlas Antibodies - 1000 NEW Products

2007-12-03T10:18:00.001-08:00

1,000 NEW HPA Antibodies
More than 700 immunohistochemically (IHC) stained tissue and cell sample images available for each HPA Antibody
Immunofluorescence (IF) staining images available for more than 800 HPA Antibodies
More than 1,000 Western Blot (WB) images available

for more information please visit us at: www.atlasantibodies.com

Quick Blocking for Perfect Westerns

2007-11-30T01:09:00.001-08:00

When the milk gets old, it's time to throw it away. Our 5M QuickBlock Kit can completely block any solid surface in just five minutes, reaching the same level of antigen detection with smaller amounts of antibody. http://www.biologyproject.net/quick_block_kit.html · Quick procedure: Efficiently blocks a western blot or ELISA plate in only five minutes. · Increased sensitivity: Significantly increases the ratio of signal over background noise, thus requires much less amount of antibody. · Wide range: Works with over 80% of the antibodies from the most popular species · Easy optimization: Determines the best blocking reagents for problematic antibodies · Low cost: The kits are designed only for blocking. There are no bundled reagents to inflate the price and depress your budget. Our customer representatives are available 24 hours, Monday through Friday. You may contact us anytime for assistance. Please check our contact page http://www.biologyproject.net/contact.html for our ocal reginal numbers.Western Blot Specialist Gen Script Corporation – VWR Strategic Partner 120 Centennial Ave., Piscataway, NJ 08854 Tel: 1-732-885-9188 ext 128 Fax: 1-732-210-0262

Custom Peptide Synthesis

2007-11-30T01:07:00.000-08:00

Custom Peptide Synthesis

With almost 10 years experience in solid phase peptide synthesis, SBS Genetech is committed to supplying high quality peptides at competitive prices and providing our customers with the extensive services and products:• Purities from crude to 98% • Acetylation/Amidation • Phosphorylation/Fluorescein/Biotin labeled peptides • Cyclic peptides • KLH/BSA Conjugation • MAP peptides• 2-3 weeks turn-around time.

Check the price here
http://www.biocompare.com/optin/20071129_2077.html

Purification of miRNA from FFPE Sections

2007-11-28T10:23:00.001-08:00

Purification of miRNA from FFPE Sections

The miRNeasy FFPE Kit from QIAGEN enables purification of total RNA that includes miRNA and other small RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections including laser capture microscopy samples. Fixing tissues with formalin leads to RNA–RNA and RNA–protein crosslinking, which impairs RNA performance in downstream applications. The miRNeasy FFPE Kit reverses formalin crosslinking and releases RNA from tissue sections while avoiding further RNA degradation. Optimized binding conditions allow purification of all usable RNA from approximately 18 nucleotides upwards. The purified RNA delivers maximal performance in a range of applications, including real-time PCR. To find out more about the miRNeasy FFPE Kit and QIAGEN's comprehensive range of miRNA research technologies, please visit the link below.

miRNeasy FFPE Kit

New online tool for LC method transfer

2007-11-28T10:16:00.001-08:00

Better LC results, faster than ever.
Calculate it for yourself and compare
The Agilent 1200 Series Rapid Resolution LC (RRLC) system lets you keep
all your options open. With just one system you can push RRLC
performance to new limits and continue to run your conventional LC
methods, too.

Agilent's new online tool for LC method transfer
and cost calculation lets you make your own calculations and immediately
see the results:

Transfer your conventional LC methods
to RRLC methods
Compare equipment, consumables and
labor costs for conventional LC to RRLC
Try out the new Agilent LC method translator
plus cost calculator tool www.agilent.com/chem/RRLC_MT

Nanoparticle siRNA Transfection System

2007-11-28T10:13:00.001-08:00

N-TER™ Nanoparticle siRNA Transfection System: siRNA Delivery to Difficult-to-Transfect Cells

Traditional lipid-based siRNA transfection reagents have a number of drawbacks, including a limited ability to transfect into a variety of cell types, such as primary, neuronal, differentiated, and non-dividing cells. Sigma's N-TER™ Nanoparticle siRNA Transfection System is a peptide based transfection reagent specifically designed to bypass these limitations and allow for efficient delivery of siRNAs into historically unapproachable eukaryotic cell lines. For more information, please visit us online at the link below.
more...

Novel Luciferase Assay System for HTS Applications

2007-11-28T10:08:00.001-08:00

Targeting Systems Offers a Novel Luciferase Assay System for HTS Applications
This product is based on gaussia luciferase, a secreted luciferase that is more than a 1000 times brighter than firefly or renilla luciferases. When it comes to screening compounds that affect the activity of a weak promoter, an assay type and readout method can make the difference between actually finding compounds or proteins of interest and not finding them. Gaussia luciferase is extremely attractive for drug screening involving weak promoters. Ease of use, elimination of cell lysis, signal stability, and high signal intensity make the Secreted Gaussia Luciferase System a powerful tool for high-throughput applications. Targeting Systems also offers a dual luciferase reporter system based on gaussia luciferase with firefly luciferase as the second reporter.
more...

Reproducibility in Protein Biomarker Research WEBINAR

2007-11-27T21:00:00.001-08:00

Reproducibility in Protein Biomarker Research WEBINAR

Please join our panel of experts on 5 December 2007 for a live online
educational seminar, "Tackling Reproducibility Issues in Mass
Spectrometry-based Biomarker Discovery."

Submit your questions LIVE to the experts during the webinar!

Date: Wednesday, December 5, 2007
Time: 12:00 noon EST; 9:00 am PST; 5 pm GMT
Duration: 1 hour

www.sciencemag.org/webinar

Mass spectrometry is an extremely powerful and sensitive technology that
can detect very small changes in expression levels. Some of these
changes may stem from the biological differences related to a disease or
treatment of interest. Others, however, may reflect the heterogeneity of
patients across multiple sites, the inherent biological complexity and
diversity of different sample types, and even small differences in
sample collection, processing, handling, and analysis techniques used by
multiple operators across multiple locations. As a consequence, data may
be tainted by site-, study-, population-, or sample-specific anomalies
and, therefore, not be sufficiently robust for biomarker discovery.

Join our panel of experts in a live, audience-driven Q&A as they discuss
how to overcome these reproducibility issues to generate less biased and
more reliable results. Questions for the panel can be submitted live
through your viewing console.

Participants:

Martin Latterich, Ph.D.
Faculty of Pharmacy
University of Montreal
Montreal, Canada

Timothy D. Veenstra, Ph.D.
SAIC-Frederick, Inc.
National Cancer Institute at Frederick
Frederick , MD

Toni Whistler, Ph.D.
Chronic Viral Diseases Branch
Centers for Disease Control and Prevention
Atlanta, GA
REGISTER NOW: www.sciencemag.org/webinar

--- Produced by the Science /AAAS Business Office and sponsored by
Bio-Rad ---

2007-2008 Sigma Antibodies Catalog

2007-11-27T20:27:00.000-08:00

Introducing the 2007-2008 Sigma Antibodies Catalog
This new 320-page catalog includes:
* Over 4,000 high-quality antibodies
* Monoclonals, polyclonals, secondaries & conjugates
* Over 750 new antibodies
* Application photos
* Associated gene symbols
* Technical appendices
Click here to request your free copy online at
http://www.sigma.com/stkeabcatalog

BULL'S-EYE-InvitrogenT GPCR research tools

2007-11-27T05:37:00.000-08:00

LESS MOVING TARGET, MORE BULL'S-EYE-Invitrogen(tm) GPCR research tools

Gain a clear advantage when interrogating difficult receptors with
integrated GPCR cellular assays from Invitrogen. The expert tools needed
to see the most lucrative targets, including target assays with multiple
readout options, are right in front of you. Learn more and sign up for
the Drug Discovery E-newsletter at www.invitrogen.com/gpcr.

2008 AAI Annual Meeting Alert

2007-11-01T10:54:00.001-07:00

2008 AAI Annual Meeting Alert

Only 6 days to Abstract Deadline!

Only 13 days to AAI Awards Deadline!

EB 2008 Abstracts
Due Wednesday, November 7!
Click here to submit

AAI Award Nominations, Applications
Due Wednesday, November 14!
Click here for details

FOR 2008!
AAI Trainee Abstract Awards!

Due Wednesday, November 7!

AAI Annual Meeting

Preliminary Program

The American Association of Immunologists (AAI)
9650 Rockville Pike * Bethesda, MD 20814
301-634-7178 * infoaai@aai.org

analyze gene expression or DNA methylation

2007-10-24T06:29:00.001-07:00

Do you knock down genes with siRNA, analyze gene expression or DNA methylation, or study miRNAs? We understand the major challenges that have to be overcome.

Our sample and assay technologies ensure that you obtain the reliable, high-quality data you need. From sample collection and stabilization to analyte purification and analysis - we’ve got it covered.

For more details, visit our application pages:

You can also find out more in our new brochure Analyzing Gene Expression and Regulation.

PCR.ppt- Microsoft PowerPoint

2007-10-22T14:25:00.001-07:00

[PDF]

Microsoft PowerPoint - PCR.ppt

File Format: PDF/Adobe Acrobat - View as HTML
Nucleotides. Primers. polymerase. PCR is DNA replication in a test tube ... Reverse trancriptase PCR. Use mRNA as a template. Isolated cDNA clones ...
learn.sdstate.edu/chend/Courses/2006.Fall/Math592.Bioinformatics/PCR.pdf -

ARK-Genomics conference 2008: 3rd International Symposiumon Animal Functional Genomics (ISAFG)

2007-10-19T02:05:00.001-07:00

ARK-Genomics conference 2008: 3rd International Symposium on Animal Functional Genomics (ISAFG)

Dates:
April 7th – 9th 2008

Venue:
Edinburgh International Conference Centre (EICC), Morrison Street, Edinburgh, UK

Official Conference Website:
http://ebrc-launch.org/ISAFG/index.html
(updated information about agenda, registration, abstract submission, accommodation and venue)

Registration and Abstract Submission will open on Friday, October 12^th 2007
Reduced rates for PhD and first year post docs available.
Limited number of travel bursaries for young researchers available.

Highlights:
The 3rd International Symposium on Animal Functional Genomics (ISAFG) will present opportunities to take stock of the current European and International position in animal functional genomics, identifying areas for collaborations and the development of new resources.

Increased exposure through the bringing together of two important conference series:
1st European Farm Animal Functional Genomics Conference (ARK-Genomics) and
2nd International Symposium on Animal Functional Genomics (Michigan State University)

Themes:
The conference will be structured around the following themes:
“Genetics and QTLs”

“Bioinformatics and Data Mining”

“Animal Health”

“Functional Genomics meets Physiology”

“Animal Genomics in Industry”

“Proteomics”

“Systems Biology”

The ISAFG event will run in parallel with the launch of the EBRC, a new world class animal bioscience centre.
The keynote lectures by Professor John Quackenbush and Professor Michel Georges will highlight functional genomics and its impact on Systems Biology.

Additionally there will be 2 open sessions, a poster session with cheese &wine as well as a conference dinner.
As before, there will be plenty of opportunities for networking.

More information about ARK-Genomics, the collaborative functional genomics research centre can be found at:

http://www.ark-genomics.org/

RNA In Situ Hybridization Protocol

2007-10-17T20:09:00.001-07:00

96-well RNA In Situ Hybridization Protocol - http://www.fruitfly.org/about/methods/RNAinsitu.html

96-well RNA In Situ Hybridization Protocol. Probe Preparation, Cell Inoculation, PCR, RNA Probe Preparation, etc, Very In-depth Protocol and Method Conditions. Berkeley Drosophila Genome Project.

http://www.fruitfly.org/about/methods/RNAinsitu.html

Milestones in DNA Technologies

2007-10-17T12:02:00.001-07:00

Milestones in DNA Technologies

A collaboration from Nature, Nature Methods and Nature Reviews Genetics, Milestones in DNA Technologies will focus on ground-breaking technologies and advances in the analysis of DNA. It will contain articles from the participating journals and original commentaries written by personalities who have witnessed the unfolding, or consequences, of these milestone achievements.

Free print copies available! Click here to request your free print copy.

Climate Scientists, Gore Share Nobel Peace Prize

2007-10-14T03:59:00.001-07:00

Climate Scientists, Gore Share Nobel Peace Prize
Committee honors efforts to spread global awareness of climate change: http://sciencenow.sciencemag.org/cgi/content/full/2007/1012/1?etoc

Neuron SFN Satellite Meeting: Register Now!

2007-10-12T09:37:00.001-07:00

Only 21 days until the meeting... Register to attend and Submit your Abstract today!

Neurons on the Map (just preceding the SfN meeting)
November 1, 2007
San Diego, CA

For the complete meeting details, visit http://www.neuron-meeting.com.

If you have any questions regarding registration, please contact Charlotte Wilkins c.wilkins@elsevier.com.

Viral Metagenomics WEBINAR

2007-10-12T09:22:00.001-07:00

Viral Metagenomics WEBINAR

Join an online seminar on 24 October 2007 entitled "A Viral Metagenomics Study of Honey Bee Colony Collapse Disorder" presented by two eminent authors of a recently published Science paper.

Submit your questions LIVE to the experts during the webinar!

Date: Wednesday, October 24, 2007
Time: 12:00 noon EDT; 9:00 a.m. PDT ; 5 p.m. GMT
Duration: 1 hour

Register now!
For more information and complimentary registration visit:
www.sciencemag.org/webinar

Colony collapse disorder (CCD) among honey bee populations in the United States has resulted in the loss of between 50% and 90% of hive colonies. Previous studies have pointed to the possibility that an infectious agent could be involved. A recent study published in Science magazine used unbiased metagenomic analysis to survey microflora present in normal and CCD-affected hives to determine whether a pathological agent could be linked to CCD. The authors found that the presence of one virus, Israeli acute paralysis virus of bees (IAPV), showed a strong correlation with colony collapse disorder. In addition to the important economical implications, this work also represents a novel use for massively parallel next generation sequencing technology which has enabled this type of high level metagenomic study.

You will hear our panel, which includes two of the study's authors, discussing:
- how metagenomics can be applied in the discovery of unknown pathogens
- the importance of study design and data analysis in metagenomics research
- how recent technological advances have made this type of study possible

Participants:
W. Ian Lipkin, M.D.
Professor of Epidemiology and
Professor of Neurology and Pathology
Mailman School of Public Health
Columbia University
New York , NY

Michael Egholm, Ph.D.
Vice President of Research & Development
454 Life Sciences
Branford , CT

REGISTER NOW AT: www.sciencemag.org/webinar

Buy EVOcard to Receive Christmas presents

2007-10-10T11:18:00.001-07:00


Buy your EVOcard and receive 2 Christmas presents on orders placed until 24th December for oligos & sequencing

Up to 35% discount off the list price on your MWG orders (35% discount on unmodified oligonucleotides; 15% discount on sequencing reactions and modified oligonucleotides) Amazon vouchers or EVOcard bonus (7.5% of total order spend until 24/12/07 arwarded as Amazon vouchers; 15% of total order spend from until 24/12/07 applied to account as EVOcard bonus) Please register by email to be eligible for the bonus - send contact details and your preference of EVOcard bonus or Amazon vouchers to evocardpromotion@mwgdna.com or faxback this postcard to + 49 80928289970.

Millipore Cell Signaling Application Guide

2007-10-10T11:14:00.001-07:00

Invaluable, up-to-date information on one of the most critical life science workflows—Cell Signaling—is now available.
Are you looking for the latest products and technical information in cell signaling? With more than 500 pages of products, workflows and technical reference material the New Cell Signaling Application Guide provides the most comprehensive support and advanced selection of tools available in the area of cell signaling.

The Cell Signaling Application Guide includes dozens of cell signaling pathways, over 100 pages of technical articles, an extensive troubleshooting section, as well as thousands of Upstate cell signaling products including detailed product information in the following areas:

• Upstate enzymes, antibodies and kits
• Epigenetics and Chromatin Function
• Multiplexing and Drug Discovery Services
• Pathway Diagrams

Request your copy of the Cell Signaling Application Guide.

Real-time PCR Video for GM soybean DNA

2007-10-10T08:27:00.001-07:00

PCR Animation

2007-10-10T08:19:00.001-07:00

PCR Tutorial Animation

Molecular Structure Animation

2007-10-10T08:15:00.001-07:00

Interleukin-1 binding to its receptor on a cell surface, created from structural data

SDS-PAGE VIDEO

2007-10-10T08:12:00.001-07:00

Biochemical Applications 101 - Sodium dodecylsuplhate gelelectrophoresis (SDS-PAGE)

Video SDS PAGE

2007-10-10T08:07:00.001-07:00

A video showing SDS polyacrylamide gel electrophoresis in ~120x time (constant voltage). Note the stained markers on the right.