Biochemistry 2280A Lecture Notes - Lecture 23: Contig, Genetic Code, Northern Blot
14 Apr 2020
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Document Summary
A collection of cloned dna fragments that represent all of the dna in an organism"s genome. Lots of human dna is isolated from many cells. Cleaved to partial digest by restriction nucleases - produces millions of fragments. The fragments are inserted into plasmids using ligase. These plasmids are introduced into bacteria --> leads to millions of clones: influenza was the first genome sequenced. Gram negative bacteria with a circular genome of 2 million base pairs. If you were to use independent clones with an average insert size of 2000 bp, you would need 1000 clones as the bare minimum - however you want a several fold to be sure. Start with millions of cells, extract dna, sonicate into dna fragments of various sizes. Purify the dna fragments and isolate fragments around 2000 bp (not too big but still a workable size) Prepare a clone library - 20, 000 clones each representing a fragment of the genome.
