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Old 01-13-2019, 11:57 PM   #15
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,232
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I can think of following two approaches:

1- Their enzymatic fragmentation leaves blunt end or/and 5 overhang which can be repaired and A tailed by Taq.

2- Mix of proofreading and non-proofreading polymerases with exonuclease resistant A nucleotide. In this case 3 overhang can be excised by proofreading polymerase, and 5 overhang can be filled in by both and A tailed by non-proofreading polymerase. Template 5 phosphorylation can be done by including PNK in ligation enzyme mix or at fragmentation step.
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