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Lysis Buffer Issues

Lysis Buffer Issues - Chemistry Forum

Lysis Buffer Issues - Chemistry Forum. Discuss chemical reactions, chemistry.


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Old 08-04-2010, 03:50 PM
Pipette Filler
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Default Lysis Buffer Issues



Hi, everyone! I am trying to make lysis buffer and when I add the DTT the buffer turns yellow. It's done this twice now. Is it possible that my DTT is bad? There's no way our bottle is more than 4 years old (kept in powder form in a -20 automatic defrost freezer). The only things I had added so far were 1M NaF, 0.5M EGTA, 0.1M Na3VO4, and about 800ml of dd H20. Any idea what's causing the yellowing? Which chemical is most likely to be bad? Thanks!!

Cynthia
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Old 08-09-2010, 03:36 PM
Pipette Filler
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Default Re: Lysis Buffer Issues

Hi, guys - I have discovered that I was making my Na3VO4 incorrectly. I didn' t know I was supposed to pH it to 10 and the boil until clear. I have now done this step correctly but the buffer *still* turns yellow! Am I just supposed to make the lysis buffer all up and not worry about it being yellow? Or am I supposed to mix it all up and then boil it until it's clear? Please help . . .
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