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lecture_notes:mar31 [2010/04/01 10:21] svasili |
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__Sequencing by synthesis/Pyrosequencing__\\ | __Sequencing by synthesis/Pyrosequencing__\\ | ||
- | Involves synthesizing complementary strand of DNA to the template DNA strand to be sequenced. Four enzymes are used in this method - DNA polymerase, ATP sulfyrylase, Luciferase, and Apyrase. The light generated is captured as a signal which detects the base incorporated. Currently the read length is approximately 400 nucleotides.\\ | + | Involves synthesizing complementary strand of DNA to the template DNA strand to be sequenced. Four enzymes are used in this method - DNA polymerase, ATP sulfyrylase, Luciferase, and Apyrase. The light generated is captured as a signal which detects the base incorporated. Currently the read length is approximately 400 nucleotides. Incorporation of homopolymer A is found to be problematic.\\ |
Further description of how the 4 enzymes work in this process can be found here[[http://en.wikipedia.org/wiki/Pyrosequencing]].\\ | Further description of how the 4 enzymes work in this process can be found here[[http://en.wikipedia.org/wiki/Pyrosequencing]].\\ | ||
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* The chemistry of Illumina sequencing platform can be found at [[http://www.illumina.com/technology/sequencing_technology.ilmn]].\\ | * The chemistry of Illumina sequencing platform can be found at [[http://www.illumina.com/technology/sequencing_technology.ilmn]].\\ | ||
- | * Now Sequencing Platform (CMOS based sequencing platform) - Sequencing based on detection of charge when a base is incorporated. Link to the article published by Nader in this respect is [[http://www.pnas.org/content/103/17/6466]]. On an average this sequencing technology incorporates upto 2.5 nucleotides in 2 minutes and in 60 minutes upto 100 bases.\\ | + | * Now Sequencing Platform (CMOS based sequencing platform) - Sequencing based on detection of charge when a base is incorporated. Link to the article published by Nader in this respect is [[http://www.pnas.org/content/103/17/6466]]. On an average this sequencing technology incorporates upto 2.5 nucleotides in 2 minutes and in 60 minutes upto 100 bases. No problem is detected so far with the incorporation of homopolymer A.\\ |
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- | Also we meet in the science library this Friday. | + | Also we meet in the science library (instructions room near periodicals) this Friday. |