I understand the reasons why too much starting template can inhibit the formation of PCR amplicons, and I am aware of the guideline amounts of DNA in weight/volume measurements. But obviously the same amount of copy numbers will not be present in a 50ug/ml DNA sample extracted from a virus that has a genome size of 100,000 base pairs compared with the same concentration of DNA extracted from a plant that has a genome size of 150Gb. So what is the optimal starting DNA template copy number desired for a PCR reaction (assuming of course that the primers will only bind the targeted region of the genome?)
this is a recommed dosage from my pfu pCR kit (50ul)
genome DNA: 100-200ng
plasmid DNA: 1-10ng
cDNA: 50-100ng
λDNA: 1-10ng. hope this will help you.
Well, as per our experience it can be around 25 to 50 copies. Real time PCR can detect these copies in a 40 cycles amplification program, Ct would be around 38-39.
Approximately 104 copies of target DNA are required to detect product in 25-30 PCR cycles
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