Insert size?

Przemko przemko at reks.uia.ac.be
Tue Nov 23 05:00:30 EST 1993


Hi out there!
I have a question that will probably prove that I slept on my
undergrad. Anyway, here it goes. Why it is so difficult
(down to impossible) to clone large fragments of DNA into plasmid
vectors. Practically, anything above 10kb is very difficult
to clone. If, on the other hand one would use a phage (or even
better: a cosmid) the problem seems to be gone. In fact it becomes
quite opposite: small fragments are difficult to clone. My problem
is that ColE1, the grandaddy of all plasmids is about 100kb long.
So, the size alone is not an issue. Is it topology, or something
particular about replication or what?
I post it here hoping that this is the right group. There are so
many of them these days (Sigh)...Bionet this, bionet that, each of
them with one post a week. Anyway
ThanX for the answers
Przemko Tylzanowski Ph.D.
Dept. Biochemistry
University of Antwerp

It was posted on sci.bio but apparently the quastion is not
scientific enough, so maybe someone who is ACTUALLY doing
the bench work will help me



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