In-gel electroporation?

[Viraj A. Master]

In article <3g3l50$al0 at lyra.csx.cam.ac.uk>, cdw1000 at mole.bio.cam.ac.uk
(Christopher Walentas (Bioc)) wrote:

| Flipping through the FMC booklet that comes with NuSeive,
| I saw they had a protocol for in-gel ligations and 
| transformations.  Is it possible to use electroporation
| with a gel slice as well or due to the electrophoretic
| buffers (salt) is one stuck the Hanahan and the like?
| Any advice on the subject that you could forward to me
| (incl. other sources to chase down) would be appreciated.
| 
| Cheers in advance,
| 
| Tripp


Hi Tripp,


I do in-gel ligations according to the protocol of 

Kalvakolanu and Livingston (1991) Biotechniques 10:176 

and then use 4microliters of the diluted ligation (1:1 with water) for
electroporation (using 80microliters of cells).  It works well.

Good luck,

Viraj

-- 
Viraj Master
Department of Organismal Biology and Anatomy
University of Chicago