Plasmid loss after restreaking
ddh at unlinfo.unl.edu
Tue Jul 18 09:04:46 EST 1995
I have a problem which I think was discussed in detail very recently.
Could any one post the conclusion of that discussion?
"PROVERB IS NOT A PROVERB UNLESS YOU EXPERIENCE IT"
My problem: I transformed Sure18 cells with recombinant pBluescript
plasmid. Selected the white transformants on LB+Amp+X-gal+IPTG plates.
I did a master plate of half a dozen transformants. I was unable to
extract plasmid DNA using mini-prep method. And now the cells wouldn't
grow either in plates (Amp) or in LB medium supplemented with Amp. I
guess I have lost the plasmid.
Now: 1. Should I retransform in different cell type like DH5-alpha or
XL1-Blue? Does it help in any way?
2. Or I should think of using X-gal dissolved in a different
Thanks in advance.
More information about the Methods