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8 Cards in this Set
- Front
- Back
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Next Gen Sequencing - 4 steps |
Do not rely on chain termination 1) template strand is immobilized 2)amplified into large number of identical fragments 3)reagents added 4)sequenced by synthesis -hundreds/thousands strands are sequenced in parallel |
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Main advantages of NGS over Sanger (5) |
-Cost - human genome sequenced for 5K -Sample Size - less template required -Speed - massively paralleled -Accuracy - sequenced multiple times -not limited to organisms with published genomes |
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NGS includes |
a number of diff modern sequencing techs (6 in slide) |
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Roche 454 Pyrosequencing |
**need clarification -fragments are attached to bead -added to well - the light signal released is correlated with the number of nucleotides joined to the growing strand (ex: GG is brighter than G) |
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Life tech ion torrent sequencing |
**need clarification -DNA is fragmented -added to bead -amplifyed by PCR -placed in single well of slide -slide is flooded with dNTPs, releases H ions, decreases pH -pH is detected |
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Illumina Sequencing |
**Need clarification slide is flooded with flouroscently labelled nucleotides -image is taken of each "cycle" -fluorescent signal indicates which base has been added
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5 applications of NGS |
1. Agricultural Genomics -improve drought tolerance of corn hybrids 2. Cancer Genomics -sequence normal tissue, to identify mutations in tumour tissue 3. Forensic Genomics -disaster victim identification, complete info from highly damaged samples 4. Complex disease genomics -not all diseases are associated with one gene - detect genetic variants to multiple samples 5. Microbial Genomics -sequencing bacteria |
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International Cancer Genome Consortium (ICGC) |
What: large scale caner genome study for 50 diff cancer types Rationale: -Scope: huge, no country can do it all -Reduce duplication -Standardization and Uniform Quality -Accelerate dissemination |
