PCR and knockout

My point is that the primer doesn't come off after replication, so PCR reaction is actually limited by the amount of nucleotides and primers that are added.

Knock-out using transposon, EZ-Tn5 in my case, is entirely random. But knock-out is done in vitro with PCR-ed gene of interest. The knocked out gene piece of DNA is then put back into the bacteria for homologous recombination.
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