Hi there,
I'm trying to grasp how the nick translation works exactly after ligating the P1 and P2 adapters to A-tailed DNA fragments. Does the reaction at 72oC exchange the nucleotides of the insert DNA (5' to 3') or does it go in the other direction, only exchanging the nucleotides in the adapter sequences towards the 'outisde'? This is very important to us since we want to bisulfite-convert our libraries after ligation. Can anyone shed light on this?
I'm trying to grasp how the nick translation works exactly after ligating the P1 and P2 adapters to A-tailed DNA fragments. Does the reaction at 72oC exchange the nucleotides of the insert DNA (5' to 3') or does it go in the other direction, only exchanging the nucleotides in the adapter sequences towards the 'outisde'? This is very important to us since we want to bisulfite-convert our libraries after ligation. Can anyone shed light on this?
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