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Direct Sequencing (Sanger Sequencing) Method
Services • Methods

Direct Sequencing (Sanger Sequencing) Method

A method of direct reading of the DNA sequence of all coding exons and exonic/intronic boundaries of a particular gene, associated with the disease. In this method, the nucleotide sequence of a DNA fragment is determined using automatic sequencers. The separation of the DNA fragments is performed using capillary electrophoresis. Each of the four nucleotides is marked with four different fluorescent dyes, which, after laser irradiation, emit light of different wavelengths. This light subsequently is detected by a camera and transformed by specialized software into an electropherogram. The sensitivity of Sanger Sequencing is 99,5%.

Sanger sequencing is used for carrier testing of an already identified pathogenic mutation in the family and/or for confirmation of a variant, detected with Next Generation Sequencing technology.

LGD offers the following molecular diagnostic tests using the direct sequencing method: