GENETICS - Investigations (PCR, FISH, Linkage, Blotting) Flashcards

1
Q

What is the optimal temperature for DNA polymerase?

A

70 degrees

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
1
Q

What is the optimal length for primers in PCR?

A

17-20 base pairs

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Which molecular techniques are mutation detection techniques?

A
  1. DNA sequencing
  2. Southern blotting
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

What type of technique is used in Restriction Fragment Length Polymorphisms (RFLPs)?

A

Southern Blotting

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What are the steps involved in the cloning process of DNA? (4)

A
  1. Restriction endonucleases cleave sections of human DNA
  2. DNA ligase incorporates human DNA into a vector
  3. Vector transfers DNA to bacteria
  4. Bacteria replicate required segment of DNA
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What is the difference between southern blotting and northern blotting?

A

Southern - DNA sequences
Northern - mRNA sequences

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

What is the main limitation of PCR?

A

High false-positive rate

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What is the difference between
southern blotting
northern blotting
western blotting
south-western blotting?

A

southern blotting - detects DNA sequences
northern blotting- detects mRNA sequences
western blotting-detects specific proteins
south-western - DNA-bound proteins (transcription factors)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

What are the main reagents needed in a PCR? (6)

A
  1. DNA - sequence to be copied
  2. Taq polymerase - builds new strands
  3. free nucleotides - building blocks
  4. Primers - short sequences that adhere to start of target region - tell taq polymerase where to start building from
  5. Buffers - maintain pH/concentration of ions for optimal conditions
  6. Mg2+ cofactors - primer adheres at correct site and helps taq polymerases to work correctly
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What are the steps in PCR? (3)

A
  1. Denaturation - boil to 95 degrees to break DNA strands to create two template strands (breaks hydrogen bonds)
  2. Annealing - 55 degrees allows primers (18-22 nucleotides in length) to stick to template areas of DNA
  3. Extension - 72 degrees - synthesis of complementary DNA by TAQ polymerase synthesising from 5’ to 3’ direction

Cycle is repeated 50 times, DNA is amplified 105 times.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What is FISH and what is it used for?

A

FISH is a method of visualising specific locations on a chromosomes - allows detection of chromosomal abnormalities using flourescent DNA probes.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly