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Created on: 15-May-2014. Updated on: 17-May-2014.

Category: Nucleic Acids.

Colony PCR (screening of successful ligations)

Posted by: dami82, UIC, Chicago, IL, http://www.biotechworld.it/portfolio/fantini



This procedure is based on PCR and allows the screening of a high number of single transformant colonies (E.coli) for the presence (and/or the length) of a specific DNA sequence. Since part of the PCR reaction mixture is inoculated in LB, transformant E.coli corresponding to the successful PCR reactions can be further grown and used for plasmid preparation.
It is important to carefully design the colony PCR experiment before starting (i.e. include appropriate positive and negative controls and analyze enough colonies).

Prepare a PCR cocktail according to the following scheme (per samp...

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