resuspended primers in water - is this a problem?! (Sep/09/2008 )
My PI ordered some primer stocks. I resuspended them in water instead of TE. How bad is that? Is there anything I can do?
Water is even better than TE buffer. I always resuspend my primers in PCR-grade water or better in low Tris buffer (10mM Tris, pH 8.0) - try to avoid TE because the EDTA in the buffer chelates a part of the MgCl2 in the following PCR reaction.... If you do not want to store them in water, add a small portion of concentrated Tris buffer and correct the concentration accordingly
I agree. I also use for my primers. Store them at -20°C or -80° and you will be fine.
Thank you guys so much! I really appreciate it.
TE doesn't make any difference at all! IF you resuspend the primer at 100 uM, you still have to dilute it ~20-fold to get to your working concentration (at least we do). Any EDTA would be diluted to 50uM - hardly an inhibitory concentration.
The benefit of TE is you kill any nucleases that might get into the stock solution.
i never resuspend anything in TE, i alays use nuclease free water.. good luck