Plasmid loss after restreaking

dinakar bhattramakki ddh at unlinfo.unl.edu
Tue Jul 18 09:04:46 EST 1995


Hi netters,

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 
     organic solvent? 

Thanks in advance.
--
- Dinakar



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