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-   -   Lysis Buffer Issues (http://www.molecularstation.com/forum/chemistry-forum/73496-lysis-buffer-issues.html)

SmallRodent 08-04-2010 03:50 PM

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 :)

SmallRodent 08-09-2010 03:36 PM

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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