Classical Sequencing

Question 1

dATPalphaS is a nucleotide that is used in preference to dATP in pyrosequencing because:

Answer 1

dATP can drive the reaction to light emission even if it is not incorporated into the growing DNA chain

Question 2

What is the actual sequence read between the red demarcation lines in this autorad?

Answer 2

5’-AGGTTATCAGATCTGCAACTCATATGATCGAGAGGG-3’

Question 3

Here is a sequence autorad for a normal individual (control) and a patient; what is the difference between them?

Answer 3

C > T

Question 4

Which of the following components terminates the chain in a sequencing reaction?

Answer 4

Dideoxynucleotides

Question 5

With respect to Sanger sequencing of DNA, which of the following statements is not true?

Answer 5

To visualise the DNA, radioactivity must not be used

Question 6

All of the following are necessary for chain-termination sequencing except:

Answer 6

All are necessary

Question 7

What is the major drawback associated with this sequencing technology?

Answer 7

Correlation loss between light intensity and high number of repetitive nucleotides

Question 8

True or false: four distinct fluorophore emissions are emitted in this sequencing protocol; the emission wavelength per single cycle is dependent on which of the four nucleotides was incorporated.

Answer 8

False

Question 9

Here are two depictions of dideoxy sequencing readouts. Which readout refers to dideoxy sequencing via capillary electrophoresis?

Answer 9

B