Protocols
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Blunt End Cloning PCR Products
(HanCock Lab)
http://www.cmdr.ubc.ca/bobh/methods/BLUNTENDCLONIN...
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Cloning of PCR Products with Phosphorylated Primers
(Hideaki Shiraishi, Kyoto University)
For cloning DNA fragments by ligating with dephosphorylated vector DNA, the fragments should have phosphates on their 5' termini. Since phosphorylation of PCR products by T4 polynucleotide kinase is inefficient, primers should be phosphorylated prior to PCR reaction.
http://kuchem.kyoto-u.ac.jp/seika/shiraishi/protoc...
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Cloning PCR Product
(Fermentas)
Including PCR primer design, DNA preparation, ligation, transformation...
http://www.fermentas.com/Profiles/Kits/PDovetailKi...
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Cloning PCR-Generated DNA Fragments
(Gerard Lazo)
http://wheat.pw.usda.gov/~lazo/methods/lazo/pcr-cl...
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Direct Cloning of Blunt-end PCR Fragments
(Gerard Lazo)
http://wheat.pw.usda.gov/~lazo/methods/lazo/met4.h...
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One Tube PCR Cloning Method
(Chun-Ming Liu)
This method has 3 reactions (restriction, polishing and ligation) carried out in one tube using ligation buffer. Believe or not, it works very well without using expensive kits.
http://www.plant.dlo.nl/projects/hybtech/Liu/onetu...
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One-Tube PCR Cloning
(Hideaki Shiraishi, Kyoto University)
In the following protocol, recombinant plasmids can be generated in a single reaction tube from an intact plasmid and unpolished PCR products. Although this procedure is simple and rapid, number of colonies obtained after transformation of E. coli is usually smaller than that obtained by standard cloning techniques.
http://kuchem.kyoto-u.ac.jp/seika/shiraishi/protoc...
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PCR Product Cloning
(Michael Blaber's Lab)
This is a very useful guide to PCR cloning. Three strategies are described in great detail including introducing a restriction site, generation of half site and TA cloning.
http://wine1.sb.fsu.edu/bch5425/lect24/lect24.htm
Added: Tue May 14 2002, Hits: 1306, Reviews: 0 Write review Cached