Posted by: sYaKuuRiolAKU_nchImb September 28, 2011
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Shiva nagar is i guess right,
40 degrees is too low for a 20 mer primer with workable proportion of GC and AT
also depends on what enzyme you are using, Taq, pfu or phusion., kappa etc phusion requires more Tm and pfu less. In either case 40 is low
you are expected to get
1. primer binding to itself
2. more non specific product
3. no product at all
read the answers above from other users if you are looking for a basic PCR reaction mechanism
40 degrees is too low for a 20 mer primer with workable proportion of GC and AT
also depends on what enzyme you are using, Taq, pfu or phusion., kappa etc phusion requires more Tm and pfu less. In either case 40 is low
you are expected to get
1. primer binding to itself
2. more non specific product
3. no product at all
read the answers above from other users if you are looking for a basic PCR reaction mechanism