Reason for odd PCR conditions
Posted 29 December 2010 - 09:21 AM
I am amplifying off of plasmid DNA a 600 bp product. I am curious as to why I have one set of steps (repeated 15x) with an elongation of 30 sec and then another set of steps (repeated 25x) with an elongation of 45 sec. I am using Taq polymerase. The PCR works great, IO just don't understand the need for 2 sets of steps.
Posted 30 December 2010 - 08:22 AM
longer elongation time just for synthesize longer DNA fragment~
May be this is a special technique (although i dont think so). Let see how other ppl answer
Posted 30 December 2010 - 11:02 AM
Posted 30 December 2010 - 11:56 AM
This is e.g. used in cross-species amplifications (primers of different species are tried out for a new one).
Anyway perhaps you overlooked the temperature decrease?
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.