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Tips on how to plasmid rescue p{PZ}?

Gregg Roman roman at bcm.tmc.edu
Fri Jun 25 07:58:33 EST 1999


Dear Ze,

        Have you performed Southern blots to map the predicted size of the
rescue?  If your flanking genomic fragment is too large you may be better
off trying inverse PCR rather than optimizing for the rescue of a 30 kb
plasmid.   iPCR is quick, and you have several enzyme choices available for
obtaining flanking genomic DNA.  Also, don't forget that you can use AvrII
in combination with XbaI to rescue PZ elements.  Good Luck!

Gregg Roman
Department of Cell Biology
Baylor College of Medicine
Houston,  TX  77030
.
ze cheng wrote in message <19990624002958.17825.qmail at hotmail.com>...
>Hi,
>    I am a graduate student of SUNYAB.My research project involves plasmid
>rescueing p{PZ} from p-element containing stains.I tried several times in
>vain so far,even with the combination of digestion with XbaI and NheI or
>XbaI with SpeI.The problem here is that I have to clone a very big
>plasmid(at least 7Kb).Has anyone succeeded in doing that?Can someone give
me
>some tips on how to ligate and transform effiently?Thank you very much.
>   My address is:ze_cheng at hotmail.com
>   Does anyone know how to subscribe to the newsgroup?
>   ginger
>
>
>_______________________________________________________________
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