Precipitate in PCR buffer w/BSA & 2-mercaptoethanol - PCR buffers (Oct/03/2006 )
Good morning. I was given a recipe for a "special" PCR buffer for GC-rich sequences, which contains Tris.HCl, pH 8.8, ammonium sulfate, BSA 10mg/mL, and 100mM 2-mercaptoethanol. I made the buffer, but when I went to use a tube (the lab stores 10x buffer stocks frozen), all of them had a white precipitate! Someone else had a problem with another buffer containing BSA and dithiothretol, so she now adds DTT separately. Is there any way to add both BSA and 2-mercaptoethanol to the stock solution, so I do not have to pipet ridiculously tiny amounts, and avoid the precipitate?
You can probably redissolve the precipitate before use by heating the buffer to 37 degrees for a few minutes and then vortexing well.
But if the only problem is you have to add less than 1 ul, simply dilute your DTT 10 fold so you can add an amount that is easy to pipet.