Designing PCR primers-How to check them?

SEHuang sehuang at aol.com
Sun Jul 23 18:44:30 EST 1995


Using rather longer primers (around 30 mer) and high annealing temps
(around 70 degree C, you may even use one temp step for anneal AND
elongation) i have recently performed more than ten different PCR's with
different primer pairs on genomic DNA without bothering about self
priming, primer dimer and non-specific annealing to genomic targets - all
primer pairs worked well at first try. Depending on the protocol
(especially when ampifying PURIFIED DNA) hot start may be necessary.
Good luck

S. Huang
Children's Hospital
Boston



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