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Genome sequencing. Haixu Tang School of Informatics. Methodology for DNA Sequencing. The chain termination method (Sanger et al., 1977) The chemical degradation method (Maxam and Gilbert, 1977) . Chain termination method. Polyacrylamide gel electrophoresis . Sequencing vectors. PCR.

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genome sequencing

Genome sequencing

Haixu Tang

School of Informatics

methodology for dna sequencing
Methodology for DNA Sequencing
  • The chain termination method (Sanger et al., 1977)
  • The chemical degradation method (Maxam and Gilbert, 1977)
clone vectors
Clone vectors
  • Bacteriophage P1 vectors (Sternberg, 1990) can clone larger fragments of DNA, up to 125 kb using current technology.
  • Bacterial artificial chromosomes or BACs (Shizuya et al., 1992) can be used to clone fragments of 300 kb and longer.
  • P1-derived artificial chromosomes or PACs (Ioannou et al., 1994) combine features of P1 vectors and BACs and have a capacity of up to 300 kb.
  • Fosmids (Kim et al., 1992) contain the F plasmid origin of replication and a l cos site. They are less prone to instability problems.
human genome project
Human genome project
  • 300,000 BACs  “sequence-ready” map
  • Shotgun sequencing of each BAC
  • Order BACs to get the genome sequences