Cloning PCR Product
Including PCR primer design, DNA preparation, ligation, transformation...
Added: Tue May 14 2002, Hits: 4471, Reviews:
3 of 33 people found the following review helpful:
Extra complication without adequate advantage
, Thu Nov 06 2003
Old-School(?) Molecular Geneticist
This cloning method appears to offer no substantial benefit compared with introducing 5' recognition sequences for type II restriction enzymes and digesting directly at these sites. The product literature mentions that, in cases where 2 different enzymes are employed, digestion conditions may not match. But many restriction enzymes perform wonderfully in double-digests, and NEB even provides a handy reference chart (http://www.neb.com/neb/tech/tech_resource/restriction/buffers/double_digest.html). Why bother with longer primers and less reliable type IIS restriction enzymes when simpler methods emplying type II enzymes work perfectly well?
Maybe the folks at Fermentas can clear this up?
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