In chain termination sequencing we synthesize new strands
of DNA which are complementary to a single-stranded template. The template DNA is
provided with a mixture consisting of all four deoxynucleotides with each of them
labeled with a different colored fluorescent tag and DNA polymerase. The presence of all
four deoxynucleotides allows chain elongation to proceed till dideoxynucleotide is
inserted by the DNA polymerase. The outcome of this is a new set of DNA chains all of
which have different lengths. Using gel electrophoresis the fragments are separated
based on their size. In the detector as the labeled DNA fragments pass a detector at the
bottom of the gel, the color is recorded. The different colors which represent the
various nucleotide sequences allow the DNA sequence to be reconstructed from their
pattern.
Wednesday, December 16, 2015
Describe how a chain termination sequencing reaction works, and how the products of the reaction are detected.What are the properties that the...
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