DNA Quantitation Part 1 (Exam 1)

Question 1

This is the movement of charged particles in an electric field.

Answer 1

What is electrophoresis?

Question 2

Is DNA loaded at the cathode or anode?

Answer 2

Cathode

Question 3

What sets PCR from rtPCR?

Answer 3

rtPCR uses a fluorescent DNA probe and collects information on a cycle-by-cycle basis.

Question 4

List the 5 components needed for PCR.

Answer 4

1) Template DNA
2) Taq polymerase
3) Primers
4) Buffer w/ Mg2+ ions and such
5) Deoxynucleotidetriphosphates (dNTPs)

Question 5

What are the 3 steps w/ associated temperatures (in Celsius) for PCR?

Answer 5

1) Denaturation (95)
2) Annealing (50-65)
3) Extension (72)

Question 6

What are 4 components of a primer?

Answer 6

1) Artificially synthesized
2) ssDNA
3) Complementary to target sequence
4) 20-30 bases long

Question 7

What species is rtPCR designed for and why?

Answer 7

Humans because of probe specificity

Question 8

What is the purpose of a DNA probe?

Answer 8

To target and fluoresce a certain sequence

Question 9

What is the relationship between reporter and quencher dyes on a DNA probe?

Answer 9

Quencher dye absorbs the reporter’s fluorescence until the probe is degrade by primer extension, meaning fluorescence is unbound and detectable