BIOC 4403 Study Guide - Final Guide: Satellite Dna, Helicase, Transfer Rna

• School: Dalhousie University
• Department: Biochem & Molecular Biology
• Course: BIOC 4403
• Professor: Archibald John

Document Summary

Chain terminator (sanger) sequencing: single-stranded template, primer polymerase, all four dntps (one labeled*, one of four ddntps (very low conc. ) in four separate tubes. Read from bottom to up, the sequence of complementary strand. Sanger methods achieve read lengths of approximately 800bp. remains in wide use, for smaller- scale projects, validation of next- Gen results and for obtaining especially long contiguous dna sequence reads (>500 nucleotides) Shotgun sequencing: uses fluorescently-labeled ddntps, thermal cycle sequencing reaction to incorporate the ddntps and produce labeled chain-terminated products. All four synthesis / chain termination reactions done in the same tube; Sample run in a single lane on the gel, runs past a laser beam which causes fluorescence, received by a detector, converted into a chromatogram . Faster and more reactions in parallel; the limit for the individual reaction 800bp. Impractical for molecules many kb long genomic dna is broken in large pieces and cloned into special vectors;