No way.

Paul N Hengen pnh at
Tue Aug 8 14:31:18 EST 1995

 In article <1995Aug7.201657.12045 at>
 murrayb at (Bernard Murray) writes:

| Nikolai writes:
|: Yesterday I found that the only clone had died!!! (dried out in incubator;
|: technician forget to add water). The month of hard work trashed.
|: It kills me, I need rest. I want vacation.
| Wait!!! Don't throw out that dead culture! You can transform the plasmid
| back into E. coli. Add competent cells on ice for a long time, swirl to
| keep the plasmid DNA hydrated, heat shock, express REALLY long with glucose
| in the media, plate everything on selective media, and pray like hell you
| get one transformant :-)
| OR, you can collect all dried out cells, wash with 1M NaCl, and lyse them
| according to the alkaline lysis method. Collect plasmid DNA and transform
| all of it into highly competent E. coli cells.

> Ah, this assumes that it is a *plasmid* clone!
> What if it was a cell clone?
> Hopefully it was a nice easy plasmid that could be rescued...

Sorry about that! Since it wasn't clear, I assumed it was a bacterial culture
with recombinant plasmid.  But, in fact Nikolai wrote to me that it was a cell
line....probably gone forever :-(

BTW, FWIW the plasmid thing does work. You can even transform with the media
less E. coli cells. There is a paper out there in Literature-Land about
plasmids being excreted into the media, but I can't remember where I've seen
it. If you know which one I'm talking about, please tell me. I would like to
keep track of them by adding to my database.

* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
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