MCDB Christoffersen Lecture#12

MCDB Christoffersen Lecture#12 - MCDB Christoffersen...

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MCDB Christoffersen Lecture #12 Cloning DNA Source of Insert DNA (where the DNA is inserted): genomic DNA, cDNA, PCR product DNA Vector (takes insert DNA and copies it in a host): plasmid, virus, artificial chromosome Host: E. Coil, yeast, cultured eukaryotic cells Ligate insert and vector to make recombinant DNA molecule in test tube. Transform recombinant DNA molecule (a way for host to take DNA up to host) into host which will replicate and make many copies of each clone Genomic vs. cDNA Gene Library A genomic library is a population of host bacteria, each of which carries a DNA molecule that was inserted into a cloning vector, such that the collection of cloned DNA molecules represents the entire genome of the source organism. This term also represents the collection of all of the vector molecules, each carrying a piece of the chromosomal DNA of the organism, prior to the insertion of these molecules into the host cells. Types of Vectors: o Plasmids: Have a capacity of 15 kb. o Phage (lambda)s: Have a capacity of 25 kb. o Cosmids or Fosmids: Have a capacity of 35-45 kb. o Bacterial artificial chromosomes (BAC)(P-1 derived): Have a capacity of 50-300 kb.
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This note was uploaded on 01/19/2012 for the course MCDB 1a taught by Professor Feinstein during the Fall '08 term at UCSB.

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MCDB Christoffersen Lecture#12 - MCDB Christoffersen...

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