# How to make up a buffer?? - (Sep/27/2011 )

Can any one please help explain the calculations on how to make up the following;

lysis buffer (50mM Tris-Cl pH 8, 10mM EDTA, 2% SDS).

I'm totally lost on where to begin.

So what do you know about making up buffers?

If starting from powdered chemicals, you will need the following equations V=n/C (where V =volume, n=number of moles and C=concentration) and n=m/M_{r} (where n= number of moles, m= mass and M_{r}= molar mass).

Please read the Material safety data sheet (MSDS) before using any chemicals, and use appropriate safety equipment - if you don't have them, ASK FOR THEM!

EDTA is a bit tricky - you need the correct pH before it will dissolve.

Only thing i know about making buffers is using Henderson Hasselback equation pH= pKa + log (

I'm not even sure if thats correct and if its to be made up to a volume of 100ml. I'm really lost for this question no matter how hard i try i can't come up with a logic answer.

Haras911 on Wed Oct 5 13:05:08 2011 said:

Only thing i know about making buffers is using Henderson Hasselback equation pH= pKa + log (

you don't have to use the equation to make up a buffer. just have an appropriate acid and base component and use one to adjust the other to the desired pH.

I'm not even sure if thats correct and if its to be made up to a volume of 100ml.

it would be made up to 100ml.