MGY377H1 Final: Study notes for exam
25 Dec 2011
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Sequencing procedures to determine the exact order of nucleotides in the dna fragment. Maxim and gilbert method improved throughput, see sequence from where cleaved. But sanger method is the standard currently. using primers complementary to the plasmid area flanking either end of the insert. after sequencing thousands of plasmids this way the sequences can be stitched together by a computer to assemble the full genome. Cloning bias some gene segments will be toxic to e. coli and cannot be cloned: result in gaps in the assembled sequence that must be filled with more laborious sequencing methods. Project maintenance issues: maintain a lot of different e. coli isolates. Slow and expensive: many runs need to be done. Real time sequencing get sequence as reaction progresses. Can be done in liquid or with dna substrate attached to slide or bead.
