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These platforms are classified into two :\\ | These platforms are classified into two :\\ | ||
* 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.\\ | ||
- | * Sequencing by hybridization : SOLiD.\\ | + | * Sequencing by ligation : SOLiD.\\ |
__Sequencing by synthesis/Pyrosequencing__\\ | __Sequencing by synthesis/Pyrosequencing__\\ | ||
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* 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]].\\ | * 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]].\\ | ||
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* Illumina/Solexa \\ | * Illumina/Solexa \\ | ||
- | * Randomly fragment genomic DNA and ligate adapters to both ends of the fragments | + | * Preparing DNA sample : Randomly fragment genomic DNA and ligate adapters to both ends of the fragments.\\ |
+ | * Immobilize DNA to surface : Bind single stranded fragments randomly to the inside surface of the flow cell channels.\\ | ||
+ | * Bridge amplification.\\ | ||
+ | * Sequence Colonies : In first chemistry cycle determine first base. Get rid of dye, otherwise chances of mis-incorporation.\\ | ||
+ | * Sequence reads over multiple chemistry cycles.\\ | ||
+ | * Read length not fixed.\\ | ||
+ | * The chemistry of Illumina sequencing plaform can be found at [[http://www.illumina.com/technology/sequencing_technology.ilmn]].\\ | ||
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+ | * 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.\\ | ||
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+ | __Sequencing by Ligation__\\ | ||
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