hroychow at NMSU.EDU
Thu Feb 26 12:50:42 EST 1998
At 02:31 PM 2/25/98 -0800, Xiao-Hua Xue wrote:
>Hi, dear netters,
> I used BclI (from GIBCO) to digest a plasmid and got some problems: 1)
>there is only very weak band showing up which is supposed to be the
>digested DNA, while undigested DNA still show high density. I assume this
>is due to incomplete digestion. 2) there is a big smear in front of that
>weak digested DNA band. I assume this is due to nonspecific digestion. I
>tried a few times by changing concentration of BclI, digestion time (from
>2hr to overnight), at temperature 50oC and results are similar.
> Does anybody have experience on BclI digestion. I really appreciate if
>you could give me some suggestions. Thanks in advance.
What host are you getting your plasmid from? BclI site is methylated in most
commonly used hosts. Try getting your plasmid from GM33 or a similar dcm-
Dr. Hiranya Sankar Roychowdhury
Plant Genetic Engineering Lab.
New Mexico State University
Las Cruces, NM 88003
Ph. (505) 646-5785
hroychow at nmsu.edu
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