Biochemistry 2280A Lecture Notes - Lecture 27: Southern Blot, Arginine, Consensus Sequence
23 Apr 2016
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Craig venter and francis collins used independent approaches and pooled their data. Traditional genome sequencing involves: creating a genomic dna library, many independent sequencing reactions, aligning the independent sequences into a continuous sequence. Genomic dna library: a collection of cloned dna fragments that represent all of the genomic dna (this includes non-coding regions, regulatory regions, etc. ; protein coding genes can even be split in half between different clones because restriction fragments do not distinguish this) in an organism. Reverse transcriptase is then added to create a ss cdna copy (primer is poly-t tail), and dna polymerase next to create ds cdna. These cdna fragments are then introduced into plasmid vectors and amplified in bacteria. However, a cdna library will only contain protein coding genes, and the specific orf of a gene without introns, where as a genomic library will contain everything (regulatory sequences, introns, exons, etc).
