Slide 2:
Sponge Dry Tissue Papers Weight Southern Blotting
(Southern, 1975) Gel DNA Nitrocellulose
membrane Animated S-Blot
Slide 3:
Probing / Hybridization Animated S-Blot
Slide 4:
How do we get polymorphism?
Substitution leads to RFLP………………… AGCTTATTCGGATTCAAGGATCCTTCGGATTCAACTA MUTATED
G to T AGCTTATTCGTATTCAAGGATCCTTCGG ATTCAACTA RESTRICTION
FRAGMENTS AGCTTATTCGGTTTCAAGGATCCTTCGGATTCAACTA Results into
Restriction Fragment Length
POLYMOPRPHISM deletion
insertion and/or
transposition
or error in DNA replication
ACT IN THE SAME WAY © A.K. Chhabra
Slide 5:
RFLP markers arise as a result of
Mutations:
substitution of a single nucleotide
rearrangements in the DNA intervening between
two restriction sires Such rearrangements
might include
deletion
insertion and/or transposition
or error in DNA replication. © A.K. Chhabra
Slide 6:
How
Substitution leads to RFLP………………… AGCTTATTCGGATTCAAGGATCCTTCGGATTCAACTA MUTATED
G to T AGCTTATTCGTATTCAAGGATCCTTCGG ATTCAACTA RESTRICTION
FRAGMENTS AGCTTATTCGGTTTCAAGGATCCTTCGGATTCAACTA Results into
Restriction Fragment Length
POLYMOPRPHISM deletion
insertion and/or
transposition
or error in DNA replication
ACT IN THE SAME WAY © A.K. Chhabra
Slide 7:
RFLP Technique
DNA isolation (text file)
Digestion of the DNA with a restriction enzyme
Separation of the restricted fragments by agarose gel electrophoresis/PAGE
Southern Blotting
Detection of individual restriction fragments
by nucleic acid hybridization
with labeled cloned probes,
Scoring of RFLPs by direct observation of autoradiograms. © A.K. Chhabra RFLP ANIMATED