Hi,
We are hoping to carry out NGS of PCR amplified DNA aptamers (80 bp). We have PCR’d up the DNA aptamers using Taq pol (which should generate a 3’ A overhang) and then treated using PNK (to generate a 5’ phosphate). As I understand it the DNA is now in the correct configuration for adaptor ligation (using the Tru Seq PCR free kit), is this correct? The alternative is to end-repair the PCR products and A-tail, but is this necessary if I use the Taq pol – PNK method as above?
Any advice much appreciated
C
We are hoping to carry out NGS of PCR amplified DNA aptamers (80 bp). We have PCR’d up the DNA aptamers using Taq pol (which should generate a 3’ A overhang) and then treated using PNK (to generate a 5’ phosphate). As I understand it the DNA is now in the correct configuration for adaptor ligation (using the Tru Seq PCR free kit), is this correct? The alternative is to end-repair the PCR products and A-tail, but is this necessary if I use the Taq pol – PNK method as above?
Any advice much appreciated
C
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