Dideoxysequencing, also known as Sanger sequencing, is a method used to determine the sequence of nucleotides in a DNA molecule. The method involves using a DNA polymerase enzyme, DNA primers, fluorescently labeled dideoxynucleotides, and a template DNA strand.
The DNA polymerase enzyme extends a DNA primer by adding complementary nucleotides to the template strand, and when a dideoxynucleotide is incorporated into the growing DNA strand, DNA synthesis stops. This results in the creation of fragments of different lengths, each terminating with a dideoxynucleotide. The fragments are then separated by size using gel electrophoresis, and the sequence is determined by analyzing the pattern of fluorescent signals generated by the different colored dideoxynucleotides incorporated into the fragments.
This method was developed by Frederick Sanger in the 1970s and is widely used in molecular biology research and genetic analysis.
what is dideoxysequencing method
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