I'm using the ready to go PCR beads for the first time. The protocol recommends a final volume of 25 uL. I tried using 2 uL of primer mix (1 uL of each primer, at a concentration of 5 uM, diluted from a 100 uM stock), 100 ng of template genomic DNA, and diluted to 25 uL. so far I have not gotten clear bands. Is there anything in particular when using the beads that would help with the results?
http://www.gelifesciences.co.jp/tech_support/manual/pdf/27955701pl.pdf
I've always had great luck with the beads even for degraded DNA. Here's what I add to the tube:
1 ul of forward primer at 10 uM
1 ul of reverse primer at 10 uM
3 ul of DNA
20 ul sterile water
If you are still having problems, maybe there is a problem with the PCR program or the primers are not appropriate.
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