Genomic libraries are a collection of clones: low qualities of re will partially digest dna, creates overlapping fragments, library contains complete genome, probe for any gene of interest cdna libraries are created from mrna, mrna must be reverse transcribed to cdna, reverse transcriptase, reverse transcribes rna to dna. Made by retroviruses dttt primer allows for total mrna to be reverse transcribed cdna libraries cdna inserted in plasmid: like total dna total cdna inserted into plasmid, cdna lacks introns. Pcr: producing dna by cloning is time intensive, multiple steps, bacteria, polymerase chain reaction is much quicker, in vitro, take small amount of dna, specifically amplify dna of interest, essentially replication of a portion of a dna molecule. Pcr background: second revolution: in genetics, first is restriction enzymes, based on thermo cycling, naturation, annealing, elongation. Ordered clone sequencing: know vector sequence, start sequencing from vector, create probe to end of fragment, pull out new clone with match sequence, sequence new clone, repeat.
