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BIOL 102 (194)

Biotechnology cont'd (identification, genomic libraries, PCR, etc)

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Queen's University
BIOL 102
Daniel Lefebvre

Don’t need to know ch 16. Bit of 17 Restriction Enzymes • Base pairs usually palindrome-like • First letter from genus + first 2 letters from species + (first letter from strain) nomenclature • Cloning using restriction enzyme: - cut DNA into fragments with short single-stranded regions called sticky ends - sticky ends hydrogen bond with each other - DNA ligase closes gaps Recombinant DNA • Contains DNA from 2+ sources (not necessarily from different species) DNA library • Collection of many recombinant vectors each with a fragment of DNA derived from an organism o Genomic – inserts derived from chromosomal DNA o cDNA – makes DNA from mRNA of interest (uses enzyme calledreverse transciptase); lacks introns so simpler to use Genomic Library • plasma  cell: digest chromosomal DNA with a restriction enzyme and ligate pieces into vectors • transform bacteria with recombinant vectors; vectors also carry a gene that confers resistance to ampicillin • put on petri dishes containing ampicillin and allow cells to grow and divide Identifying Gene Families • Southern blotting o can detect presence of a particular gene within a mix of many chromosomal fragments separated on a gel • Possible through Electrophoresis o separates macromolecules based on charge and size (negative DNA move towards positive electrode, and smaller fragments move fast/farther)  tutorial • Possible also through Southern blotting of a gel o probe b
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