Low Efficiency from Electroporation

Esbjorn Fiers esfie at gengenp.rug.ac.be
Thu Nov 26 07:21:46 EST 1998


Hi Susan,

I have been doing electroporation for about 2 months in XL-1 Blue without any
result (low efficiency and almost no colonies from the ligation). I made new
competent cells though they didn't work neither! So I decided to go on using
others: DH5alfa. Now everything worked out fine. So if it is only for cloning
purposes and you r not bound to those XL-1 blues, try an other strain!
Makes sure the smear on your gel is not from RNA, if you use the birnboim
isolation method you should add some RNase to your loading dye before adding
it to your DNA preparations.
Well I could give you a lot more tips, though I don't know whether they would
be appropriate so when you need more information contact me via e-mail with
more details about your problems.
Hope this is useful for u,

Esbjorn Fiers


--
==================================================================
Esbjorn Fiers
DEPARTMENT OF GENETICS                         Fax:32 (0)9 2645349
UNIVERSITY OF GENT, K. L. Ledeganckstraat 35, B-9000 Gent, Belgium
Vlaams Instituut voor Biotechnologie                           VIB
mailto:esfie at gengenp.rug.ac.be        http://spider.rug.ac.be
==================================================================





More information about the Methods mailing list