Sanger and his colleagues were the first to sequence an entire genome of bacteriophage (viral DNA) which contained 5286 base pairs.
By: Nida, Sharon and Selene
How does the Sanger Dideoxy Method Work?
1.) The DNA template to be sequenced is treated so that it becomes single stranded.
4.) DNA replication begins in each
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The Human Genome Project
The Human Genome Project (HGP):
is an international scientific
research project with the goal of determining the sequence of chemical base pairs which make up human DNA,
and of identifying and mapping all of the genes of the human genome from
both a physical and functional standpoint.
Real World Applications
known as dideoxy sequencing or the Sanger ..
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The Maxam Gilbert Method
The Maxam-Gilbert method differs from the Sanger-dideoxy method in that it involves the degradation of the DNA chain while the Sanger-dideoxy method involves the synthesis of a complementary chain.
allow DNA chain synthesis to occur through a condensation reaction ..
There are 4 different dideoxyribonucleoside triphosphate chain-terminating molecules (, , , ) each containing one of the DNA bases, so ddCTP contains the base ‘C’ () etc, complementary to a on the template strand. These 4 different molecules are used in 4 separate DNA synthesis reactions on copies of the same single-stranded DNA template strand to be sequenced. When a ddNTP is incorporated into the growing newly synthesised chain, it terminates synthesis at that point so each reaction produces fragments of different lengths complementary to the template strand, which have terminated at different points.
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A DNA primer binds to the 5' end of the single-stranded DNA molecule to be sequenced (the template strand) to allow the DNA polymerase enzyme to attach and synthesize a new DNA chain against the template strand through Watson-Crick complementary base pairing.
In Sanger sequencing, the target DNA is copied many ..
ddNTP lacks the -OH group on the 3' carbon of the ribose sugar, and thus if incorporated intoa nucleic acid chain during DNA synthesis, terminates synthesis, producing DNA chains of varying length, that all terminate with the same 3' base.
DNA sequencing | Learn Science at Scitable
DNA sequencing by radioactive labelling is thankfully no longer necessary due to the advent of dye-terminator chemistry, whereby a single strand of DNA can be labelled with a fluorescent dye corresponding to the base at the 3'end of the fragment.