Thank you both.
Except for the 12 bases at the 5 end side where I have my restriction site, the rest of the primers anneal to the target sequence.
I'm trying one more round with the Kapa HiFi strictly following their technical datasheet info. For the long primers which have the Ta above 68 deg they recommend two stage PCR setup but in my case it doesn't work.
If this last attempt doesn't work I'll switch to Pwo and see what happens.
volver
Member Since 10 Jan 2013Offline Last Active Jan 13 2013 10:53 AM





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