Vector gets shorter when insert is added.
Harry.Witchel at Bristol.ac.Uk
Thu May 15 07:28:55 EST 1997
Greetings fellow molecular explorers!
In explicably, when ligating a 3.5 kb insert (65% GC rich, with
a veriety of hairpin structures) into pcDNA 1.1 AMP (InVitrogen, 4.8
kb), I got colonies which produced plasmids with the insert, in which
the vector was only 3.5 kb. In the original gel where I purified the
vector (EcoRI digested, heat inactivated, CIAP treated, heat
inactivated, NuSieve 1% gel purified, agarase purified), the vector is
the correct size. In mapping the minipreps, the insert seems to be
normal and full length. I have found the insert in different colonies
in different directions. My bug is XL1 blue MRF'.
Anyone else get anything like this? Any explanantions?
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