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Old 12-19-2018, 02:49 PM   #11
cmbetts
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Any polymerase with 3' Exonuclease activity can do end repair in the presence of nucleotides, which would include many PCR polymerases. Here's a paper using Pfu for old school cloning https://academic.oup.com/nar/article...rectedFrom=PDF. Now heat inactivating them to do subsequent A-tailing would a tall order as the DNA will denature long before the enzyme is killed (and if you don't remove it, it will chew off any new nucleotides you'd want to add). That's why you more typically see a mesophilic enzyme used to polish, which is then heat killed automagically when you A-tail with Taq.
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