There are two sequencing-by-synthesis methods (pyrosequencing and Illumina rever

Question

There are two sequencing-by-synthesis methods (pyrosequencing and Illumina reversible terminator methods) as well as the older chain termination method with fluorescently-tagged dideoxynucleotide terminators (Sanger sequencing). During the individual sequencing reaction & data collection for these methods, there are fundamental differences in expectations because of the methods’ properties, although the data analysis will produce the same final DNA sequence for the template if all goes well. Compare and contrast in depth (evaluate) the three methods (pyro, Illumina, and Sanger) for the biochemical components/reaction in template sequence determination (at the nucleotide level) and detection of that result. Do not just write an explanation of the three techniques. Point out what you find similar or different and explain those aspects (could be one, two or all three methods showing the similarities or differences).

For example: In the two sequence-by-synthesis methods, the oligonucleotide primer that is used as the starting point for the polymerization reaction in sequencing must be complementary to a sequence that was added as an adaptor to the template DNA because the template is generated from sheared DNAs and is itself generally unknown. In contrast, because the templates in Sanger sequencing are clones or PCR products, the sequencing primers are designed from prior sequence information about the input template DNA.

Explanation / Answer

Differences:

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