VIII
Contents
Chapter 4
Electrophoresis of DNA and RNA Fragments
FENG QIAN AND GREGORY G. GERMINO • . . . • • • . • • . • • . . • • • • • • • 71
Subprotocol1: The Recovery of DNA Fragments ................ 75
Subprotocol2: Electrophoresis of RNA ....................... 77
Chapter 5
Single-Strand Conformation Polymorphism (SSCP) Analysis
FENG QIAN AND GREGORY G. GERMINO . . . • • . . . • . • • . • • . . . • • • • 83
Chapter 6
Denaturing Gradient Gel Electrophoresis
KAI-OLAF NETZER • . . • • . • • • • • • . . • . . • • . • • • • . • . . • . • . • . . . . . . 86
Chapter 7
Introduction of Pulsed-Field Gel Electrophoresis (PFGE)
FENG QIAN AND GREGORY G. GERMINO • • . • . • • . • . . • . • • • • • . . . . 96
Chapter 8
Pulsed-Field Gel Electrophoresis: Protocols
GUDRUN A. RAPPOLD, KARIN RIED, ALBRECHT KLINK,
ERCOLE RAO AND BIRGIT WEISS ...........••.•..•.•.•••.•.• 103
Subprotocol 1: Preparation and Processing of High Molecular
DNA in Agarose Blocks ...................... 104
Subprotocol 2: Preparation of Size Markers .................... 106
Subprotocol 3: Restrietion Enzyme Digestion of DNA
in Agarose Blocks . . . . . . . . . . . . . . . . . . . . . . . . . . 108
Subprotocol 4: Gel Electrophoresis .......................... 110
Chapter 9
Radioactive Labeling of DNA and RNA Probes
PETER IGARASHI .••..................•..••.•.•.•.••..•.. 114
Subprotocol1: End-Labeling Using T4 Polynucleotide Kinase ...... 114
Subprotocol2: End-Labeling Using the Exchange Kinase Reaction .. 116
Subprotocol 3: Labeling 3' Ends with Terminal Transferase
(Forward Reaction) . . . . . . . . . . . . . . . . . . . . . . . . . 117
Subprotocol4: Random-Prime Labeling ...................... 118
Subprotocol 5: Riboprobes ................................. 121