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PCR Based Cloning Kits: Something For Everybody

Date: April 13, 1998PCR Based Cloning Kits Table The End Table (PDF Format) PCR has found applications in almost every imaginable facet of molecular biology, and for many applications, looking at a band on a gel is not enough. Sequencing, expressing, mutating--all require cloning. And as it happens, cloning strategies that work for other types of DNA fragments don't work at all well, or require inordinate effort, with PCR fragments. For example, the most commonly used cloning strategy requires

Laura Defrancesco

Date: April 13, 1998PCR Based Cloning Kits Table
The End Table (PDF Format)
PCR has found applications in almost every imaginable facet of molecular biology, and for many applications, looking at a band on a gel is not enough. Sequencing, expressing, mutating--all require cloning. And as it happens, cloning strategies that work for other types of DNA fragments don't work at all well, or require inordinate effort, with PCR fragments. For example, the most commonly used cloning strategy requires that the insert and vector have compatible ends. Thus, in order to clone PCR fragments by this technique, restriction enzyme recognition sites have to be engineered into the PCR primers, adding to the time required and cost of cloning. Another common cloning strategy employed, blunt-end ligation, particularly useful when working with incompatible ends, is inefficient when applied to PCR fragments because many polymerases used in PCR, as it happens, add...

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