BIOL309 Study Guide - Midterm Guide: Lambda Phage, Genomic Library, Electroporation
16 Mar 2013
School
Department
Course
Professor
Document Summary
Competency: the ability to take up dna by transformation: coli can be induced to take up dna by: To improve transformation efficiency, use specialized host strains that have mutations in: endonucleases, recombination genes. Bacteriophage: virus that infects bacterial host and exists as a parasite. Bacteriophage lambda has linear dna genome when in capsid. Lytic: infects, multiplies, and kills host cell by lysis: insertion vectors: 7. 6 kb inserts, replacement vectors: 22 kb inserts, cosmids: plasmid with cost site -> 32-45 kb of inserts. Dna copies of mrnas -> used for overexpression, reporter fusion, etc. Genomic library: collection of clones containing all fragments of particular genome. Creating a genomic library: choose vector, fragment genome into pieces -> small to fit into vector, ligate vector and genomic dnas -> transform into host cells, evaluate quality of library, store library. Short digestion time, suboptimal mg2+, low enzyme concentration, and can fractionate. Partial digests: used to recover fragments adjacent to specific fragment.
