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How to make up a buffer??

Lysis Buffer

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#1 Haras911

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Posted 27 September 2011 - 12:04 PM

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.

#2 bob1

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Posted 27 September 2011 - 04:18 PM

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/Mr (where n= number of moles, m= mass and Mr= 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.

#3 Haras911

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Posted 05 October 2011 - 05:05 AM

Only thing i know about making buffers is using Henderson Hasselback equation pH= pKa + log ( [A-] /[HA ) but just can't get my head around it.
Im guessing the 2% SDS is made up like this- 2g of SDS dissolved in 100ml of dH20.
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.

#4 mdfenko

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Posted 05 October 2011 - 05:49 AM

Only thing i know about making buffers is using Henderson Hasselback equation pH= pKa + log ( [A-] /[HA ) but just can't get my head around it.

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.

Im guessing the 2% SDS is made up like this- 2g of SDS dissolved in 100ml of dH20.
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.
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