Biochemistry 2280A Lecture Notes - Radiography, Xeroderma Pigmentosum, Rodenticide
12 Dec 2012
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11/30/2012: creating a genomic dna library, many independent sequencing reactions, aligning the independent sequences into a continuous sequence. Collection of cloned dna fragments that represent all of the dna in an organism"s genome. Each cloned dna is like a different book in a library. Together all the clones, represent all the dna in the genome. Constructing a genomic library: genomic human dna is partially cleaved with restriction nuclease to obtain millions of genomic dna fragments, dna fragments are then inserted into plasmids using ligase and introduced into bacteria. Genome about 2 million base pairs. Q: the h. influenza genome is 2 million base pairs. A: 1000 clones (2 000 000 base pairs / 2000 base pairs per clone) would be absolute minimum. Sequence the ends of the genomic clones: one clone isolate plasmid dna, anneal primer, and obtain dideoxy sequence.
