dideoxy sequencing

dideoxy sequencing
(= Sanger dideoxy sequencing)
The most popular method of DNA sequence determination (cf. Maxam-Gilbert method). Starting with single-stranded template DNA, a short complementary primer is annealed, and extended by a DNA polymerase. The reaction is split into 4 tubes (called "A, C, G or T") each containing a low concentration of the indicated dideoxy-nucleotide, in addition to the normal deoxynucleotides. Dideoxynucleotides, once incorporated, block further chain extension, and so each tube accumulates a mixture of chains of lengths determined by the template sequence. The 4 reactions are denatured and run out on an acrylamide sequencing gel in neighbouring lanes, and the sequence read up the gel according to the order of the bands.

Dictionary of molecular biology. 2004.

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