Molecular biology techniques


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Unformatted text preview: te a DNA fragment containing the full length cDNA coding for a gene ORF cDNA a) Use a clone from cDNA library that happen to contain already the whole sequence (fished for example by hybridization, or present in a library that has already separated into individual clones that have been sequenced) b) Use sequences from database (e.g. partial sequences from cDNA libraries, and/or sequences from genome) to design primers for PCR. c) Take 2‐3 fragments containing overlapping parts of ORF from a cDNA library, and reconstitute the ORF by DNA cloning. Full length cDNA for a particular gene are the main tool to construct recombinant DNA for expression, and to construct various mutants (see examples here below) 5. Aim: Source of DNA: Steps: Examples of applications: Expression in bacteria Produce large amounts of the prot...
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