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lecture_notes:mar31 [2010/04/01 03:44] jstjohn |
lecture_notes:mar31 [2010/04/01 08:48] svasili |
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These platforms are classified into two :\\ | These platforms are classified into two :\\ | ||
- | 1) Sequencing by synthesis (SBS) : 454, Illumina/Solexa, Helicos, Pacbio, Charge based detection system Now Sequencing.\\ | + | * Sequencing by synthesis (SBS) : 454, Illumina/Solexa, Helicos, Pacbio, Charge based detection system Now Sequencing.\\ |
- | 2) Sequencing by hybridization : SOLiD.\\ | + | * Sequencing by hybridization : SOLiD.\\ |
- | __Sequencing by synthesis__\\ | + | __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. 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.\\ |
+ | Further description of how the 4 enzymes work in this process can be found here[[http://en.wikipedia.org/wiki/Pyrosequencing]].\\ | ||
+ | |||
+ | * 454 - Small ~20micron beads capturing DNA loaded onto PicoTiter plate along with the required enzymes for sequencing. The chemistry of the 454 sequencing can be found in detail at [[http://454.com/products-solutions/how-it-works/sequencing-chemistry.asp]].\\ | ||
+ | * Illumina/Solexa \\ | ||
+ | * Randomly fragment genomic DNA and ligate adapters to both ends of the fragments | ||
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