Re: cloning help!# Biology - 生物学
y*e
1 楼
If contamination goes 99.9% in all clone, that means you get a supercoiled
circular DNA as your contaminant. It transforms at a much higher efficiency.
First thing you might do is change plasmid.
I talked it to my boss, he had an interesting theory. If your insert confers
toxicity to E.coli, he said there might be some wierd recombination event that
cut the insert out of the plasmid, along with flanking reagions (which might
contain the MCS, that is why you cannot cut it again with your REs).
circular DNA as your contaminant. It transforms at a much higher efficiency.
First thing you might do is change plasmid.
I talked it to my boss, he had an interesting theory. If your insert confers
toxicity to E.coli, he said there might be some wierd recombination event that
cut the insert out of the plasmid, along with flanking reagions (which might
contain the MCS, that is why you cannot cut it again with your REs).