RFLP

Question 1

do we need a PCR?

Answer 1

• no
• instead: restriction enzyme

Question 2

what does RFLP stand for?

Answer 2

Restriction Fragment Length Polymorphism

Question 3

where do the restriction enzymes come from?

Answer 3

• Brestriced enzymes isolated from bacteria

Question 4

what to the enzymes do?

Answer 4

• detect restriction sites
• cuts DNA in short fragments (6-10bp)

Question 5

prerequisite to use this method

Answer 5

• know genome of species in advance

Question 6

what is this method used for?

Answer 6

• are individuals homozygous or heterozygous for specific alleles depending on restriction sites size

Question 7

how does it work?

Answer 7

1. detect restriction site specific of 2 alleles
2. use corresponding restriction enzyme
3. enzyme cuts DNA in fragments at that site
4. fragments of diff. lengths
5. fragments inserted in agarose gel
6. when electricity is applied: they move to pole

Question 8

Answer 8

Question 9

negative aspects

Answer 9

• time consuming
• lot of work to find right enzyme and diff. restriction sites of a target

Question 10

useful for

Answer 10

• species identification
• only if restriction sites are species specific
• useful to test, if an individual we found, is of a species with these restriction sites

Question 11

case study

Answer 11

• 2 species of sturgeons were distinguished