I am aiming to create 5 - 10 kb amplicons by PCR with all thymidines replaced by different analogs (i.e. BrdU, EdU, CldU, IdU). For this, I tried to simply replace dTTP by dBrdU (or other) in the master mix.
Having tried this for the different analogs with different polymerases (Taq, Q5, HF, Platinum Taq, Phusion HF), I haven't got any product. Whilst I found a paper that has successfully incorporated BrdU using ampliTaq (see attached) and ordered this polymerase, I am in the meantime wondering if any of you have experience with this and has any suggestion on what (not) to try.
Any suggestions would be very helpful, thanks!
Article Generation of nonradioactive bromodeoxyuridine labeled DNA p...
Correction: I did use the triphosphate versions, but thanks for this correction.
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