Colony screening

Bertrand Collet b.collet at abdn.ac.uk
Thu May 31 05:39:44 EST 2001


Hi everybody,

Several people from our group noticed a unexplained phenomenom:

When doing TA cloning, we firstly screen the colonies by PCR. Most of
the time, when we use the 2 specific PCR-primers it does not work but
when we use one plasmid specific primer plus a specific PCR-primer it
works beautifully. Is there anybody who noticed this before ? What's
behind it ?

I noticed this in PCR products from 200 bp to 2 Kb, and each time the
sequence is fine but the PCR on the colonies works only if you use 1 or
2 plasmid primer.

Thanks in advance for your explanations,

Bertrand
-- 
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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