FINDING DISEASE GENES

Question 1

Probes do not have to be __ complementary.

Answer 1

100%

Question 2

What is a cDNA probe?

Answer 2

complementary only to exons

Question 3

How is DNA prepared for gels?

Answer 3

1) DNA is first acquired
2) then cut using restriction enzymes
3) loaded onto agarose gel
4) voltage applied
5) stained

Question 4

SDS- PAGE gel

Answer 4

• nuclease digestion
• keep cold to avoid degradation
• charge affects mobility
• antibody probe used

Question 5

SOUTHERN BLOTTING used for __(a)__ analysis. This is digested prior using restriction enzymes and fractionated onto gel. The absorbent ___(b)___ performs the ‘blotting’. Membrane bathed in labelled __(c)___.

Answer 5

a) DNA
b) membrane
c) probe

Question 6

NORTHERN BLOTTING used for __(a)___ analysis. Key point is that no __(b)___ is necessary.

Answer 6

a) RNA

b) digestion

Question 7

WESTERN BLOTTING involves transferring electrophoretically fractionated __(a)__ from a gel and visualising specific __(a)__ using __(b)___ probes. Also no __(c)___ required.

Answer 7

a) proteins
b) antibody
c) digestion

Question 8

Uses of RFLP mapping?

Answer 8

1. can aim to map chromosomes
2. distinguish disease gene loci in diagnostics
3. measure genetic divergence

Question 9

Digested genomic DNA will usually appear as a continuous what?

Answer 9

smear