MCB 2050 Lecture Notes - Lecture 2: Polyacrylamide Gel Electrophoresis, Restriction Fragment Length Polymorphism, Dna Sequencing
22 Jan 2020
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Restriction mapping physical mapping of dna based on relative location of re sites. Points to remember: determine if the dna is circular or linear. If circular # if fragments = # of sites (1 re site / 1 fragment) If linear # of fragments = # of sites +1 (1 re site / 2 fragments) Two-step procedure: transfer and immobilize the dna fragments from an electrophoretic gel onto a membrane, detect the presence of specific dna fragments through molecular hybridization. Molecular hybridization annealing of a dna strand to its complementary dna strand: requires prior denaturation (separation of the two strands of the dna either by heat or chemical treatment, hybridization (renaturation) occurs when the temperature is reduced. By exposing membrane to an x-ray film, the identification of specific dna fragments can be made. The black bands indicate location of radioactive bands on blot: multiple bands = multiple genes (and long genes)
