BME80H Lecture blah

BME80H Lecture blah - 3. are engineered to contain...

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3. are engineered to contain specialized sequences (figures 13.10, 13.13, Handouts) Origin of replication (ori): allows for replication of the plasmic (rec DNA) Selectable marker (amp^R) = allows for selection of transformed cells. -usually antibiotic resistance (ampicillin res.) unique restriction site(s) -location of where insert is added in -often present in a polylinker (multiple cloning site. MCS) C. Transformation and identification of bacteria containing the recombinant DNA 1 Transforming bacteria to an ampicillia-resistant phenotype cells are transformed to an ampicillin-resistant phenotype amp^s -----amp^R amp^R 2. Selecting for bacteria-containing recombinant DNA (figure 13.12) (test on!)*pBR322: recombinant DNA-carrying cells are resistant to amp & sensitive to tetracycline (tet^R gene is disrupted by the insert) (use replica plating) puc19: recombinant DNA-carrying cells are amp^R but cannot make the product of the lacZ gene (B-gal) III working with DNA A Identifying a single clone in a genetic library 1 Genetic Libraries are a large collection of clones (recombinant DNAs) carrying different segments of DNA from a single source. Genomic libraries: libraries of all of the genes in the genome
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This note was uploaded on 01/11/2011 for the course BME 80H taught by Professor Rothwell during the Winter '07 term at UCSC.

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BME80H Lecture blah - 3. are engineered to contain...

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