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lecture_notes:04-01-2011 [2011/04/02 18:49] eyliaw |
lecture_notes:04-01-2011 [2011/04/03 19:28] (current) eyliaw |
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* Instruments: SAGE, Caliper | * Instruments: SAGE, Caliper | ||
===== Sequencing Technologies ===== | ===== Sequencing Technologies ===== | ||
- | * Sequencing by synthesis (454 and Illumina) vs sequencing by ligation (SOLiD) | + | ==== Sequencing by synthesis (454 and Illumina) ==== |
- | * Pyrosequencing (454) | + | * Synthesis limits sequencing 5' to 3' |
- | * Paired tag (Illumina and SOLiD) | + | * |
+ | ==== Sequencing by ligation (SOLiD) ==== | ||
+ | * | ||
+ | ==== Pyrosequencing (454) ==== | ||
+ | * | ||
+ | ==== Paired tag (Illumina and SOLiD) ==== | ||
+ | * | ||
+ | ==== In-house ==== | ||
+ | * Nader's lab uses a method that starts by fragmenting the DNA into long fragments (aiming for ~10k). They attach a unique barcode primer to each fragment and do circular ligation, much like SOLiD. This is followed by circular replication, primer binding, and random cleavage to obtain fragments of different lengths [(cite:circular_rep>http://en.wikipedia.org/wiki/Rolling_circle_replication)]. They sequence both ends of each fragment on the Illumina, obtaining a paired read with its 10k fragment's barcode. |