DNA Sequencing

Question 1

True or False?

If a ddA is incorporated replication will terminate

Sanger Sequencing

Answer 1

True

Question 2

The difference between WES and WGS is that…

Answer 2

WES is done on ~2% of WGS templates selected by a microband-based captur

Question 3

Massively parallel sequencing does NOT require….

aka NextGen

Answer 3

electrophoresis

Question 4

pyrosequencing

Answer 4

pyrosequencing, is founded upon the fact that each time a nucleotide is incorporated into a replicating
DNA molecule, a pyrophosphate (PPi) is released. Thus, if a succession of all four nucleotides is presented to a
DNA strand replicating on a template strand, pyrophosphate will only be released when the proper (or
complementary) nucleotide is incorporated

Question 5

Pyrosequencing does NOT require

Answer 5

dideoxynucleotides

Question 6

Sanger Sequencing with Capillary Electrophoresis

Answer 6

read as you go improvement to Sanger Sequencing

Question 7

Capillary DNA sequencing allows for simultaneous resolution of sequencing reaction products because

Answer 7

each of hte four dideoxynucleotides is labelled with a different fluorochrome that can be distinguished by virtue of the specific emission frquency

Question 8

In dideoxynucleotide sequencing adjusting the ratio to very high dATP and very low ddATP will cause …

Sanger Sequencing

Answer 8

the bands of shorter products in the A lane to fade in intensity

Question 9

A DNA polymerase would be desirable for DNA sequence analysis if it hadthe following characteristic:

Answer 9

It is highly efficient at incorporating dideoxynucleotides