RFLP

Question 1

RFLP (fingerprinting)

Answer 1

Restriction Fragment Length Polymorphism

Question 2

What is used

Answer 2

Restriction enzymes (RE) for restriction of DNA
RE examples: EcoRI, HindIII, PstI
RE are obtained from Bacteria —> bc Bacteria produce RE as a defence mechanism against viral DNA (bacteriophages)

Question 3

RFLP özellikleri

Answer 3

-based on different sizes
-different sequences can be differentiated from each other by RFLP
-RE are used to cut DNA molecules into smaller fragments
-restriction digestion, mostly used: single RE, double RE
-after restriction digestion, we run electrophoresis again to see “profiles”
-by “profiles”, we can differentiate DNA molecules

Question 4

Can RE harm host DNA

Answer 4

NO. Host DNA is protected from cleavage by its own RE through “methylation”. The host DNA is methylated at or near recognition sites, preventing the RE from cutting it.

Question 5

Ends of RE

Answer 5

Blunt ends occur when strand is cut at the same point
Sticky/protruding ends occur when strand is cut at the different points