Related Experiment Videos
Cloning and analysis of PCR-generated DNA fragments
G L Costa1, A Grafsky, M P Weiner
1Stratagene Cloning Systems, La Jolla, California 92037.
PCR Methods and Applications
|June 1, 1994
Abstract:
Methods are presented for the improved yield and analysis of blunt-ended cloning of PCR-generated DNA fragments. We show that Pfu DNA polymerase polishing of Taq DNA polymerase-generated fragments increases the yield and efficiency of cloning. Using a triple primer set consisting of two outside, asymmetrically distanced primers and one fragment-specific primer, both the presence and orientation of cloned inserts can be determined. Application of these methods allows the generation and cloning of a fragment in 1 day and the analysis of putative clones the next, thereby saving a substantial amount of both time and effort.