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lecture_notes:03-31-2010 [2010/04/02 06:12] galt |
lecture_notes:03-31-2010 [2010/04/02 18:16] jstjohn |
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**Mate-Pairing** - The chances of damage to DNA increases with time. Hence, there is a high possibility of not getting the accurate sequence data from the other end of DNA. To avoid this, the Mate-Pairing process involves the determination of sequence of DNA from the end towards the beginning and then from the beginning towards the end.\\ | **Mate-Pairing** - The chances of damage to DNA increases with time. Hence, there is a high possibility of not getting the accurate sequence data from the other end of DNA. To avoid this, the Mate-Pairing process involves the determination of sequence of DNA from the end towards the beginning and then from the beginning towards the end.\\ | ||
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- | ==== Alternative Notes ==== | ||
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- | === High Level Overview === | ||
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- | The high level sequencing workflow for all next gen tools is as follows: | ||
- | Fragment Sample -> Amplify Fragments -> Sequence Fragments | ||
- | |||
- | There are two main flavors of next generation sequencing technology: | ||
- | * Sequencing by Synthesis (SBS) | ||
- | * Hybridization | ||
- | |||
- | === Examination of Individual Technologies === | ||
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- | There are four technologies that were talked about today. | ||
- | * Pyrosequencing (454) | ||
- | * solid | ||
- | * Illumina/solexa | ||
- | * Ion Torrent | ||
== Pyrosequencing (SBS) == | == Pyrosequencing (SBS) == | ||
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== Ion Torrent (SBS) == | == Ion Torrent (SBS) == | ||
- | Works very similarly to pyrosequencing, except rather than requiring light from luciferase enzymes it takes advantage of the proton that is released when a nucleotide is added to a template. It measures this electric current. The presense of a current over a certain template when a specific nucleotide is added to solution tells you that that nucleotide was added to that template. The level of current at a template tells you the number of nucleotides that were added in that run. | + | Works very similarly to pyrosequencing, except rather than requiring light from luciferase enzymes it takes advantage of the proton that is released when a nucleotide is added to a template. It measures this electric current. The presence of a current over a certain template when a specific nucleotide is added to solution tells you that that nucleotide was added to that template. The level of current at a template tells you the number of nucleotides that were added in that run. |
Can sequence about 60 bases every 100 minutes. | Can sequence about 60 bases every 100 minutes. |