Can anyone suggest a citation to support/refute the the relationship between DNA concentrations and erroneous PCR product? I am struggling to find peer-reviewed resources that speak to this concept despite several blogs and websites presenting it as common knowledge.
Thanks!
Not from a paper, but from a Guidelines for PCR from Biolabs.
https://www.neb.com/tools-and-resources/usage-guidelines/guidelines-for-pcr-optimization-with-thermophilic-dna-polymerases
"Higher DNA concentrations decrease amplicon specificity (i.e., extra bands are more likely), particularly when a large number of cycles are employed"
In the field of HIV research, this issue has come up a lot because a typical blood sample contains thousands to millions of variants (known as a quasispecies) of HIV that are replicating in the subject's blood and lymphatic systems. It is often of interest to know the recombination rate between these variants and lineages in the subject, and therefor important to minimize or eliminate PCR-generated recombinants (partial extension of the primer on one template, melting, renanealing to a different template for further extension). Many techniques were developed for single genome amplification in order to reduce or eliminate the problem, but a key component is diluting the input template to a level where each PCR is "seeded" from a single molecule of viral template.
I do not know of a specific publication that compared results from different input template concentrations, but surely there are a few if you searched for them, not only in HIV research but in other fields.
Thanks for your help. I found a conference poster that will work for my purposes. See Figure 2A
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