BPS 3101 Lecture Notes - Lecture 5: Oligomer, Contig, Capillary Electrophoresis
29 Aug 2016
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It will hybridize to multiple clones: problem avoided if using short unique-sequence probe (oligomer) mapping close to end of clone or if pre-hybridized with repeat sequence, 2. Identify overlapping clones by finding features that they share: restriction profile fingerprinting, or clones having sts in common. Haemophilus genome project 1995 (1. 8 mbp: 1. Dna sonicated, fragments (1. 6-2kb) cloned in plasmid vectors: 2. Shotgun sequencing of insert ends of 20, 000 clones analyzed, 11 mbp of sequence, scaffolds with sequencing gaps and physical gaps: 3. Screened for overlapping clones reduced to 42 contigs: 5. Assumed gaps represented genome regions unstable for plasmid vector switched to lambda vector: 6. Probed lambda library with oligomers from contig ends or used. Pyrosequencing: one dntp is added at a time + enzyme (apyrase) that degrades dntp if not incorporated into new strand, then next dntp is added.
