about transformation of DNA into E.Coli

Fri Aug 9 10:56:52 EST 1996

On 9 Aug 1996, Sailesh Surapureddi wrote:

> 2. Initally incubate your DNA with Cells for 5min till you get  the hang
> out of your method.

How does increasing the incubation from 3 to 5 min offer any more " the
hang out of your method" 

> 3. Clean your ligation mixes, since the ligation buffers usually kill your
> cells.

If ligation buffers (2uL in 50uL cells)"usually kill" the cells, why do I 
get very high degree of transformation (about 10e8 to 10e9 transformants 
per ug of plasmid) with the control ligation (un-CIP'd blunt-end vector 
w/o the insert)? This, using the standard Hanahan methods, and ligating 
one hr at RT.


			  Hiranya S. Roychowdhury
   			  Plant Genetic Engineering Lab.
			  Box 3GL, NM State Univ.
			  Las Cruces, NM 88003
			  Phone: (505) 646-5785
			  hroychow at nmsu.edu

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