I did cloning with pGEM-T kit and transformed into a DH5a cells. I've already grown the white colonies overnight and decided to check the insert before going for plasmid extraction and sequencing. I did colony PCR using primers targeting the vector DNA flanking the insert and I got good amplification (~700bp; my insert is ~550). However, when I did colony PCR on the same samples using insert-specific primers, very few samples had amplification of the right size. So I know the samples which had amplification with insert-specific primers are reliable samples for plasmid extraction and sequencing, but should I discard those that had amplification with vector-primers but not with insert-primers?

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