rapid ligation &electroporation

Dr. Hiranya S. Roychowdhury hroychow at nmsu.edu
Fri Mar 23 20:12:51 EST 2001


Electroporation is possible because DNA molecules carry the charge across
the electrodes, as projectiles, and are essentially inserted into the cells
by the electromotive force.  Presence of too much salt causes the cations
to be the electron carriers and this causes 'arcing' across the electrodes.
The arcing kills cells. 
A microliter of the ligation mix in a minimum of about 50uL cells usually
does not cause arcing. If you are adding more than taht, you may have to
remove the salt somehow.



At 05:03 PM 3/23/01 +0100, Thomas Nilsson wrote:
>Hello,
>
>I am about to do a ligation using the Roche (previously Boehringer) rapid
>ligation kit , followed by electroporation of the ligation product. Roches
>manual states that the ligation reaction has to be purified with the "High
>Pure PCR Product purifiaction kit". Why is this? A component in the
>ligation kit interfering with the electroporation? Anybody with experience
>of electroporating rapid ligations?
>
>Thomas Nilsson
>
>
Dr. Hiranya Sankar Roychowdhury
College Asst. Prof.
Molecular Biology,
Dept. of Chemistry & Biochemistry	
Box 30001 - 3MLS
New Mexico State University
Las Cruces, NM 88003

Lab: (505) 646 4722
Office: (505) 646 8256
hroychow at nmsu.edu

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