Hello!
I´d like to know it is possible to prepare in the lab an efficient gel solubilization solution for the bands being cut in DGGE gels, to reamplify the DNA with primers with GC clamps, and if you could share any recipe it would be great.
According to a technical bulletin of Gen Elute Gel Extraction kif from Sigma, the gel solubilization solution is a final part of the process of elution of the bands of acrylamide, and is actually different from an elution solution, with which the DNA is released from the acrylamide band cut.
Thanks very much for the comments!
Martha S.
Withdraw the band properly with a new and sterile scalpel blade. Cut the gel band in half, then place it in 200µL TE buffer. Cool 30 minutes at -80°C, then warm it for 5 minutes at 90°C. Place the tube under agitation at 4°C for 2 hours. You will have enough DNA diluted in the buffer to make a PCR.
In my experience on DGGE, that dates a few years back ;-), it was sufficient to CUT the band of interest out, mince, add a small volume (eg 50 microliters) of nuclease free water and leave at 4 degrees C, ON. The following day you had enough template for PCR.. Good luck!
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