gt10 insert sequencing

Sasha Kraev bckraev at aeolus.ethz.ch
Wed Sep 28 07:02:27 EST 1994


Hi, Padmaja. If your supervisor is not against kits, you may hook one kit onto
the other in the following way
1.Get gt10 Insert Screening Amplimer Set ( Clontech 5411-1 ) and follow
the instruction. This would give you at least 1 pmol of the insert.
2.Purify this PCR product ( e.g. Qiagen Qiaquick Spin PCR Kit )
3.Get Promega fmol Sequencing Kit and follow the instruction, using primers
from the Amplimer Kit ( label with kinase, etc ). 
Yes, it is expensive, but fast and does not require much experience. You do not even have to isolate lambda DNA, because it is all done from a plaque.
As for the Silver Seq Kit, it is non-radioactive but I am afraid you will
have to play around a lot to get your 500 bp with it. Good luck

S.Kraev
bckraev at aeolus.ethz.ch

( Do I need a disclaimer? )



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