I don’t think PCR is necessarily relevant here. I had the impression that this would be lost useful at linking multi-kb fragments together. If we are looking at sizes much above 2kb, PCR is going to struggle to generate full length fragments efficiently.
replyI didn’t see this technique as having DNA modification per-se, but a novel way to managing the hybridization process. It’s stock (well engineered) oligos, if I read it correctly.
pcr amplifies all sequences, correct or wrong, no? and as I understand it, it works on short snippets the best.
replyIt amplifies sequences that contain the two primer sequences on each end of the target. So if you had synthesized sequence XYZ with some mistakes like YZX, then you could target X and Z and purify.
replyYou're correct that PCR has a limited max length, but it is longer and cheaper than vanilla DNA synthesis.
Kary B. Mullis
Nobel Prize lecture
Nobel Lecture, December 8, 1993
replyThe Polymerase Chain Reaction
https://www.nobelprize.org/prizes/chemistry/1993/mullis/lect...
Intuitively I agree some kind of selective amplification should be able to correct for the mistakes. But I think it will be complicated. Because the filtering process needs to be much more complex. It can’t just chemically match to a known subsequence - you won’t know where the mistake might be in a long sequence.
replyThis is a good point. WXYZ and WYXZ are indistinguishable via PCR. And the possibilities accumulate with more segments.
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