So, I've been doing DNA extractions and PCR of mushroom collections and cultures, and I'm having mixed results. Routinely, we've been diluting all our DNA extractions if it's above 50 ng/ul, but sometimes I'm not having good bands on gel after PCR, and I'm worried that my DNA concentration is not optimal, is there a certified optimal DNA concentration for PCR?
Curiously, I've had success with amplification of DNA templates of 0,0025 ug to 0,25 ug, but some samples with DNA amount inside this range fail to have good bands on gel, if I could know a certified dilution or DNA template mass range to work in PCR would be really nice, because sometimes I waste a whole day of work diluting and trying to confirm PCR success only to find that I didn't get any bands on gel, and I have to figure out another dilution to see if it works this time...
Hi Denis,
I am less concerned regarding the amount but rather the purity. you might further dilute the DNA to get rid of inhibitors. If you have the genome size you can easily calculate th ecopy number. Normall 1000-5000 copies as input is sufficient
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