cold-labile plasmid
Elizabeth K Winters
ekw2 at iastate.edu
Tue May 11 17:02:16 EST 1999
I am working with a plasmid which behaves abnormally at cool
temperatures. This is a 2.96 kb pBluescript vector containing a
highly-repetitive 1.6 kb cDNA insert. I have used the vector to
transform E. coli strains XL1-Blue and DH5-alpha, by heat shock and
electroporation.
When the bacteria are kept at room temperature or warmer, I can digest out
the 1.6 kb fragment as usual. However, when the bacteria containing the
plasmid are cooled to 4 degrees Centigrade or frozen (including in
glycerol stock), the plasmid digestion results in only a fragment of the
original insert.
Has anyone ever heard of a plasmid behaving in such a cold-labile way?
Do you ahve any ideas about what is happening? We suspect the fact that
the insert is highly repetitive is important.
I have more information if anyone is interested. I appreciate any input
that I may receive. Thank you.
Beth Winters
ekw2 at iastate.edu
--
Beth Winters
ekw2 at iastate.edu
"We drift down time, clutching at straws; but what good's a brick to a
drowning man?" (Tom Stoppard)
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