Orientation of a PCR product in a T-vector

Bertrand Collet b.collet at abdn.ac.uk
Fri Apr 5 07:59:06 EST 2002


Hi everyone,

I was screening colonies carrying a PCR product cloned into a T vector
by PCR, I was surprised to see that 90 % (and 100 % in another cloning)
of the colonies had the PCR product ligated in the same orientation.
I thought there was an equiprobablity of insertion in the 2 direction
for T-cloning ?
What is the rule for this (if any) ?
Thank in advance,

Bertrand

PS I ask this because I will need soon some "inverted" PCR product (the
rare orientation in the above examples) for subcloning purpose ...

--
Dr Bertrand Collet
Fish immunology group
Zoology Department
Tillydrone avenue
Aberdeen AB24 2TZ
Scotland - UK

Tel (office): (+44)(0)(1224)27 3796
Tel (lab): (+44)(0)(1224)27 2870
Fax: (+44)(0)(1224)27 2396





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