International Scholarships and Financial Aid

Sunday, 11 May 2014

How to make 1x buffer from 20x

As explained before here it is very simple and you need to understand the concept.
or you can use the formula M1V1=M2V2

Example

You have 20x Orange Juice and you wanna make 10 ml of 1x.

Simple use M1V1=M2V2

M1= Original Concentration [which is 20x]
V1= required volume to make 1x

M2= Desired Concentration [which is 1x]
V2= Desired volume which is 10 ml

20x * V1 = 1x * 10ml

V1= 0.5 ml of 20x concentrated juice is needed to add in 9.5 ml of water [10-0.5=9.5 ml]

IMPORTANT 

Don't add 0.5 ml in 10 ml. This will make final volume 10.5 ml and your concentration will be wrong. 






How to make 1x BSA


How to make 1x bsa or Bovine serum albumin (BSA)

1x BSA generally suggest that you want  make 1 gram per litre BSA solution.

You can use the following method to make
Material

Phosphate buffered

saline (PBS), pH 7.4,  10X None PS00000041 10X 50 ml 1X
Sigma-Aldrich A4919 Powder 0.5 g 1 g/L 

Preparation:

1. Weigh 0.5 g BSA and add to 300 ml purified water in a 600-ml beaker.
2. Add 50 ml of 10X PBS.
3. Adjust volume to 500 ml.
4. Store at room temperature in an appropriately labeled glass bottle.

How to make 1x buffer from 10x

This is very simple

You just need to learn the concept.

10x means 10 times concentrated

you need to make 1x

so you need to dilute it 10 time.

of you can take 1 part of 10x and mix with 9 part of water [1+9=10] to make 10x buffer.

Or you can use formula below make 1x buffer from 10x concentration buffer

M1V1=M2V2

M1 = stock concentration [10x]
V1 = volume needed of M1 concentrated stock [Z]

M2 = desired concentration [1x]
V2 = desired volume [say 10 ml you wanna make]

So as per M1V1=M2V2

10x * Zml = 1x *10ml

Z =1ml of 10x you need in 10ml of water.

This also means you need to add 10-1=9 ml of water in 1 ml of 10x concentrate to make 1x buffer.

IMPORTANT 

Don't add 1ml in to 10 ml. which would be 11 ml and your concentration will be wrong.











Saturday, 4 February 2012

How to Dilute a Stock Solution


How to Dilute a Stock Solution


A. Why dilute? 

Here are two situations that arise repeatedly in molecular biology labs:
1. You have a stock solution of some compound,  let’s say an antibiotic, and you want to add the
compound to growth medium, at a much smaller concentration than the stock solution.
2. You have a tube of very concentrated bacteria, perhaps a billion cells per milliliter. You want to put
a few hundred of them on a petri plate, so that the colonies that arise will be easily distinguishable.
In both cases, the way out of the problem is to dilute the original solution. If you work in a lab, you need to
know how to do this.

B. Methods of calculating dilutions 

1. DILUTION FACTOR METHOD (fast, but requires inspiration): First, figure out the factor by which the
original solution must be diluted. Second, divide the final volume of the desired solution by that
factor, yielding the volume required of the original solution.
EXAMPLE: Suppose you need to make a 3 ml solution of growth medium supplemented with 50 µM of
the antibiotic ampicillin from a stock solution of 5 mM ampicillin. The dilution factor is :
  (5 mM) / (50 µM) = (5000 µM) / 50 µM) = 100
 so you need to dilute:
   (3 ml) / 100 = (3000 µl) / 100 = 30 µl
 of the stock solution to a final volume of 3 ml.

Sunday, 25 December 2011

Isolation of Casein from Milk



  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.


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.




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)


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.

 

Saturday, 24 December 2011

Polony PCR Amplification


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


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

Science Protocols