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Sunday, 9 November 2014

TE buffer (Tris-EDTA buffer) Recipe

The purpose of TE buffer is to solubilize DNA or RNA, while protecting it from degradation.

10 mM Tris, bring to pH 8.0 with HCl, 1 mM EDTA

A stock can be prepared and stored at RT

Component For 500ml
1M Tris pH 8* 5ml
0.5M EDTA pH 8 1ml
dH2O 496ml

Autoclave to sterilize. It is then best to remove a working aliquot and not repeatedly access the stock.
*the pH is usually adjusted to RNA 8.0 for DNA and 7.5 for

How to make acetate-acetic acid buffer solutions

Sodium acetate-acetic acid buffer solutions, pH 3.7-5.6

Sodium acetate trihydrate, CH3COONa.3H2O, M.Wt. 136.09;
0.2M solution contains 27.22g/l.
Acetic acid, glacial is ~17.47M.

x ml 0.2M NaOAc and y ml 0.2M HOAc mixed.

pH, 18°C X mL

0.2M NaOAc

Y mL

0.2M HOAc
3.7 10.0 90.0
3.8 12.0 88.0
4.0 18.0 82.0
4.2 26.5 73.5
4.4 37.0 63.0
4.6 49.0 51.0
4.8 59.0 41.0
5.0 70.0 30.0
5.2 79.0 21.0
5.4 86.0 14.0
5.6 91.0 9.0


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.











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