Michael L. Sullivan
mlsulliv at facstaff.wisc.edu
Tue Aug 20 10:04:16 EST 2002
Make sure that when you visualize the fragments by UV, that they are
not overexposed. A long-wave hand held UV lamp is what I use for
visualizing fragments I am cutting out of gels-- this causes little
if any damage to fragments. However, many UV light boxes can destroy
fragments for cloning quite rapidly.
>Hi, my friends,
> I have sent this message once before.But till now I still cannot make it.
> Please help me.
> I'm in great trouble with my contruction of expression plasmids.The
>vectors and fragments were all double digested by the same two
>enzymes and target bands after electrophoresis were extracted out
>from gel by the gel-extraction kit
>from Roche or by freeze-thraw method.The extraction was very good
>checked by selectrophoresis.For ligation,I tried many times and
>many ways including rapid ligation kits from different companies and
>individual reagents.I can't make it always.I am so discouraged.
> Would anyone give me some suggestion and and information for my
> Thanks in advance.
> Wang Hua-zhong Ph.D
> Cytogenetics Institute,
> Department of Agronomy ,
> Nanjing Agricultural University,
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