SANGER METHOD OF DNA SEQUENCING
DNA sequence determination is based on the high resolution separation of
nucleotides on polyacarylamide or an algarose gel. One method used in determining this
sequence is called the Sanger procedure. The components used are: a polyacarylamide or
an algarose gel, DNA polymerase, a primer (a short sequence), template DNA (the DNA
to be sequenced), deoxynucleotide triphosphate, and dideoxynucleitide phosphate. With
all of these, double stranded DNA is made continuously until all the triphosphates are
used up.
The deoxynucleotides, are deoxyadenine triphosphate, deoxyguanine triphosphate,
deoxycytocine triphosphate, and deoxythymine tryphosphate. They are the same as
nucleotides with the exception being that they contain hydrogen on the 3' carbon instead
of an OH. Therefore, when these deoxynucleitides are integrated into a sequence, they
prevent the addition of further nucleotides because......
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