BIOL 200 Midterm: BIOL 200 Notes After Midterm.pdf

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17 Mar 2012
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Dr. paul berg and others were the first to successfully introduce exogenous dna fragments into bacteria to produce a recombinant gene product uprising of recombinant dna technology. Bacterial restriction enzymes cut dna at specific sites. Polylinkers are engineered with many restriction enzyme sites to facilitate insertion of dna. Ecori cuts dna at a specific palindromic sequence, gaatc. Note: vector is a specialized plasmid, which carries out specific functions upon removal of nonfunctional regions. 2 different enzymes, direction cloning results, thus it only inserts in one orientation. Dna libraries (permanent collection of genes): genomic library contain copies of dna present in the genomic/chromosomal context (intergenic regions, introns, exons, repetitive sequence, cdna library represent mrna (tissue specificity, abundance) After rt uses oligo-t primer (polydt) to initiate the ssdna synthesis that is complementary to the mrna template, rna is then removed and a polydg adapter(ggg) is annealed to the 3" end.

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