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,


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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