There is an older library prep method where samples were indexed using a 3 primer PCR method (forward and reverse primer + index primer). Illumina changed this, and I've seen people on SeqAnswers complain that it was inefficient. Unfortunately, those are the only primer types I have right now.
1) Do any labs have experience with this method, and have any tips dealing with these issues?
2) I assume that the 3 primers exist in equal amounts, but the protocol does not actually list concentrations (just volumes). Does anyone know if the primer concentrations should be equal for this amplification.
Thank you.
1) Do any labs have experience with this method, and have any tips dealing with these issues?
2) I assume that the 3 primers exist in equal amounts, but the protocol does not actually list concentrations (just volumes). Does anyone know if the primer concentrations should be equal for this amplification.
Thank you.
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