What is Sanger Dideoxy Sequencing and how is it different from Applied Biosystems Fluorescent Sequencing?
With Sanger Sequencing, DNA polymerases copy single-stranded DNA templates, by adding nucleotides to agrowing chain (extension product). Chain elongation occurs at the 3′ end of a primer, an oligonucleotide that anneals to the template. The deoxynucleotide added to the extension product is selected by base-pair matching to the template. The extension product grows by the formation of a phosphodiester bridge between the 3′-hydroxyl group at the growing end of the primer and the 5′-phosphate group of the incoming deoxynucleotide (Watson et al., 1987). The growth is in the 5′ -> Â 3′ direction (see Figure 1). DNA polymerases can also incorporate analogues of nucleotide bases. The dideoxy method of DNA sequencing developed by Sanger et al. (1977) takes advantage of this ability by using 2′,3′-dideoxynucleotides as substrates. When a dideoxynucleotide is incorporated at the 3′ end of the growing chain, chain elongation is terminated selectively at A, C, G, or T because the chain lacks a 3′-
Related Questions
- Are there problems with the default threshold setting on the Applied Biosystems 7500 when using QuantiFast Probe PCR Kits?
- What is Sanger Dideoxy Sequencing and how is it different from Applied Biosystems Fluorescent Sequencing?
- How can psychological theories can be applied to different issues in the CJS (criminal justice system)?
