Lecture 7 - Genome Analysis Methods

Question 1

What does PCR stand for?

Answer 1

Polymerase Chain Reaction (PCR)

Question 2

How do PCRs work?

Answer 2

“In vitro” synthesis of large amounts of DNA by copying from small starting quantities
Small synthetic primers (“oligonucleotides”) define the boundaries of synthesis
DNA is synthesized by a DNA “polymerase” enzyme from single “monomers” (deoxy-ribonucleotides or dNTPs)

Question 3

What are ‘the ingredients’ for performing a PCR?

Answer 3

1. Primers
2. Nucleotides
3. DNA polymerase

Question 4

What are the stages of PCR tests?

Answer 4

1. Heat denaturation 94 degrees
2. Primer annealing 55 degrees
3. Primer extension 72 degrees
4. Heat denaturation 94 degrees
5. Primer annealing and extension

Question 5

What is the most frequently used assay in molecular biology?

Answer 5

PCR applications

Question 6

What does a PCR detect?

Answer 6

Genomic sequence
Virus
Cell free circulating foetal or tumour DNA

Question 7

what else does a PCR do?

Answer 7

Generates template for other applications (Sequencing/ Other anaylsis)

Question 8

What is Sanger sequencing?

Answer 8

The sequencing technology used in the human genome mapping project

Question 9

What are the ‘ ingredients’ used for Sanger sequencing?

Answer 9

1. One primer
2. Nucleotides
3. DNA polymerase
4. Dye terminator nucleotides (ddNTPs)

Question 10

What kind of graph does a Sanger sequencing produce?

Answer 10

Output chromatogram

Question 11

Stages of Sanger sequencing?

Answer 11

Large fragments > Small fragments > Laser beam > Photomultiplier >