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L7 + L8 - PCR and DNA sequencing

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malra16's version from 2018-01-09 15:31

Section 1

Question Answer
The incorporation of a ddNMP onto a growing DNA chain stops the reaction becauseddNMP is missing an hydroxyl group on the third carbon, needed to form the phosphodiester bond
The sequence of the DNA fragments emerging from the capillary is determined bythe last base in the sequence (Fluorescent tag and elusion time)
A sequencing primer does not have to anneal directly to the gene to be sequenced ifyou clone the gene into a vector that you know, and you can sequence the whole thing.
It is preferable to label the oligonucleotide primer with a fluorescent rather than a radioactive group becausethe radioactive groups can potentially be hazardous. And the other method is just as sensitive and effective. With the fluorescent tag, the process can be automated.
Efficient covalent joining of two single-strand DNA's by DNA ligase requiresa source of energy to form the bond, and a template strand, to bring the joined into apposition.
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Section 2

Question Answer
Very long DNA fragments (>100 kb) from eukaryotic genomes can be propagated in yeast artificial chromosomes (1000 kB) and can be analyzed by chromosome walking.
Reverse transcriptase requires the following for the conversion of a single strand RNA into a double strand DNA.All four dNTP's, an RNA template and a primer.
Oligonucleotide-directed site-specific mutagnesis can be used to produce deletion, insertion and point mutation. At the same time it is useful for determining the involvement of a particular amino acid in the catalytic mechanism of an enzyme.
An example on comparative genomics is..?A study, which stated that ninety percent of human genes have counterparts in the rat genome.
Which pathway have an cellular respons to the mutated forms of superoxide dismutase?Inflammation pathway
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