M2.8. PCR & DNA sequencing

Question 1

• a method for rapidly producing multiple copies of a DNA nucleotide sequence (gene).
• allows to produce billions of copies of a specific gene in a few hours.

Answer 1

POLYMERASE CHAIN REACTION

Question 2

Requirements for PCR:

Answer 2

1. source of gene to be copied
2. Thermostable DNA polymerase
3. Deoxynucleotide triphosphates
4. A set of two oligonucleotides with complementary sequence to the gene (primers)
5. Thermostable plastic container end
6. Source of heat

Question 3

Taq polymerase, Pfu polymerase

Answer 3

Thermostable DNA polymerase:
1. derived from THERMOPHILIC BACTERIUM & THERMUS AQUATICUS
2. derived from PYROCOCCUS FURIOSUS

Question 4

dATP, dGTP, dCTP and dTTP

Answer 4

Deoxynucleotide triphosphates

Question 5

a method by which the base sequence in a DNA molecule (or a portion of it) is determined.

Answer 5

DNA sequencing

Question 6

DNA Sequencing is discovered in 1977 by?

Answer 6

Fredrick Sanger

Question 7

Selective interruption of polynucleotide synthesis using

Answer 7

2’,3’-dideoxyribonucleotide triphosphates (ddNTPs)

This interruption of synthesis leads to the formation of every possible nucleotide site mixture.

Question 8

These nucleotides are labeled using ____ during their synthesis.

Answer 8

radioactive dNTP

Question 9

The radiolabeled nucleotides are then separated on a gel by ____.

Answer 9

electrophoresis