After I did blue-white screening, I picked the white colonies and subculture on LB/Amp plate. They can grow well but when I subculture again on new plate, they cannot grow. May I know the reason? My clones are JM109 with pGEM-T Easy, the plates should be okay because I have tested with another JM109 with same plasmid, they can grow.
It sometimes happens. Some possible explanations are as follows:
1. The colonies are dead on the plate.
2. The ampicillin in the plate that your colony is on could have broken down
3. Potentially your insert is toxic to the cell if it is being expressed.
4. You could have satellite colonies (which are much smaller than actual recombinants) which are using beta lactamase produced by other cells to survive.
Let me know if that helps
I think reason 2 is not possible because my friends tried with the plate, his JM109 with pGEM-T Easy can grow nicely.
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