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Protocols
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Blunt End Cloning PCR Products
(HanCock Lab)
http://www.cmdr.ubc.ca/bobh/methods/BLUNTENDCLONIN...
Added: Tue May 14 2002, Hits: 1071, Reviews: 0 Write review Cached -
Cleavage Efficiency Close to the Termini of PCR Fragments
(Fermentas)
When designing PCR experiments in which the synthesized DNA fragment is to be subsequently digested with a RE, it is very important to determine how many extra nucleotides should be added to the 5’-end of the PCR primer next to the introduced recognition site to ensure efficient cleavage with the appropriate restriction endonuclease. Some restriction endonucleases cleave DNA poorly when their recognition sites are located at the ends of DNA fragments. Information on restriction endonuclease performance close to the end of DNA fragments may be helpful when planning the introduction of cleavage sites at DNA termini in PCR experiments.
http://www.fermentas.com/techinfo/RE/restrdigpcrii...
Added: Tue May 14 2002, Hits: 729, Reviews: 0 Write review Cached -
Digestion of PCR Products
(Fermentas)
Some basic facts you need to know to cut your PCR products.
http://www.fermentas.com/techinfo/RE/restrdigpcr.h...
Added: Tue May 14 2002, Hits: 968, Reviews: 0 Write review Cached -
Purification and Sequencing of PCR Product
(Donis-Keller lab)
This method is used to sequence directly a specific PCR product after large scale amplification. The reaction should be carried out in the usual fashion, except that after optimization of annealing temperature and other conditions in the 5 µl volume, a large scale 50 µl (10X) total reaction volume should be used to generate sufficient PCR product for sequencing.
http://humgen.wustl.edu/hdk_lab_manual/pcr/pcr3.ht...
Added: Tue May 14 2002, Hits: 6476, Reviews: 0 Write review Cached