PCR CLONING AND FASLE WHITE COLONIES

Christopher Hoover cdhoover at powergrid.electriciti.com
Wed Mar 29 18:16:22 EST 1995


Dr. Zhou:
Products of this size should clone with very high efficiency into the
TA system. Our 750 bp test product yields 80% cloning efficiency, so
you might want to PCR that control up and see if your TA Cloning
results are similar. Doing a self-ligation will also help diagnose
whether the vector has lost efficiency (it should yield 90% blue
colonies). I would also suggest checking some blue colonies in case
your product has cloned in-frame with the lacZ alpha fragment and
resulted in false negative colonies (this happens most often with small
products, 300 bp and less). You can call our Technical Support at
800-955-6288 x 2 for further assistance. 

Thanks
Christopher Hoover
Invitrogen Corp.
Technical Support



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