DNA Sequencing: Sanger Sequencing Flashcards
___ is the determination of the precise sequence of nucleotides in a section of DNA
DNA sequencing
In diagnostic laboratories, the sequencing method common ly used is ____ based on the method originally developed in 1975 by the double Nobel prize winner Frederick Sanger.
Fluorescent dideoxy chain termination
This involves a PCR - like reaction, but with just a single forward or reverse oligonucleotide primer, and utilizes a DNA template to generate a series of detectable single - stranded fragments of increasing length, each being 1 nucleotide longer than the last.
Fluorescent dideoxy chain termination
In chain termination method, a chemical reaction is first set up, which includes what five components?
DNA template; thermostable DNA polymerase
a single oligonucleotide primer
the four deoxynucleotide trisphosphate (DNTPs)
the four dideoxynucleotide trisphosphate (dDNTPs)
In chain termination method, dDNTPs are each attached to a different ___ each emitting a different wavelength or color of light.
fluorescent chemical label
During chain termination method, ____ takes place, incorporating dATP, dCTP, dGTP and dTTP, generating a new strand of DNA that is exactly complementary to the DNA template. Eventually, however, at some point, a ____ will be incorporated by chance instead of a ____ substrate molecule.
DNA polymerization; dDNTP; DNTP
In chain termination method, when dDNTP is incorporated in the chain, further elongation of that chain is halted because ddNTP lacks the ___ on the deoxyribose sugar that is necessary for the addition of the next nucleotide.
3’-OH group
In chain termination method, the PCR products are separated on the basis of ____by ____ and then electronically detected and identified by the ____upon excitation by a laser as they migrate through the gel.
length; gel electrophoresis; emitted fluorescence
