Transgenesis Techniques: Principles and Protocols Methods in Molecular Biology Vol 18

✍ Scribed by David Murphy, David A. Carter

• Publisher: Humana Press
• Year: 1993
• Tongue: English
• Leaves: 436
• Series: Methods in Molecular Biology volume 18
• Edition: 1st
• Category: Library

✦ Synopsis

Practical methods for the generation and analysis of transgenic animals, covering a comprehensive range of species, from invertebrates to lower vertebrates to higher vertebrates. Chapters examine ways in which transgenic techniques can be used to address many problems in biological systems present in-depth descriptions of analytical methods, and give step-by-step techniques that can be easily performed in your lab.

✦ Table of Contents

CHAPTER 1 --
Introduction to Transgenesis......Page 1
CHAPTER 2 -- Transgenic Rodents and the Study of the Central Nervous System
......Page 5
CHAPTER 3 -- Transgenic Animals and the Study of Cancer
......Page 21
CHAPTER 4 -- Transgenic Animals and the Study of the Immune System
......Page 35
CHAPTER 5 -- The Commercial and Agricultural Applications of Animal Transgenesis
......Page 51
CHAPTER 6 -- P Element-Mediated Germ-Line Transformation of Drosophila
......Page 70
CHAPTER 7 -- Gene Transfer by Microinjectionin the Zebra Fish Brachydanio rerio
......Page 82
CHAPTER 8 -- Transgenic Induction in Salmonid and Tilapia Fish
......Page 90
CHAPTER 9 -- An Overview of Transgenic Mouse Production
......Page 103
CHAPTER 10 -- Transgene Design
......Page 107
CHAPTER 11 -- Isolation of DNA Fragments for Microinjection
......Page 111
CHAPTER 12 -- Establishing a Colony for Efficient Production of Transgenic Mice
......Page 117
CHAPTER 13 -- Mating Mice
......Page 123
CHAPTER 14 -- Anesthetizing Mice
......Page 127
CHAPTER 15 -- Vasectomizing a Mouse
......Page 129
CHAPTER 16 -- Preparation of Culture Media for Fertilized One-Cell Mouse Eggs
......Page 133
CHAPTER 17 -- Collection of Fertilized One-Cell Mouse Eggs for Microinjection
......Page 137
CHAPTER 18 -- Microinjection of Cloned DNA Fragments into Fertilized One-Cell Mouse Eggs -- I. Manual Injection
......Page 143
CHAPTER 19 -- Microinjection of Cloned DNA Fragments into Fertilized One-Cell Mouse Eggs --
II. Automatic Injection......Page 155
CHAPTER 20 -- Delivery of Microinjected Eggs to Surrogate Mothers by Oviduct Transfer
......Page 161
CHAPTER 21 -- Caesarean Section and Fostering
......Page 169
CHAPTER 22 -- In vitro Fertilization of Mouse
......Page 171
CHAPTER 23 -- The In Vitro Isolation of Murine Embryonic Stem Cells
......Page 175
CHAPTER 24 -- Production of Chimeras Derived from Murine Embryonic Stem Cells
......Page 209
CHAPTER 25 -- Cryopreservation of Transgenic Mouse Lines
......Page 231
CHAPTER 26 -- Introduction to the Physiology and Husbandry of the Rat
......Page 236
CHAPTER 27 -- Mating of Rats
......Page 242
CHAPTER 28 -- Vasectomy of the Male Rat
......Page 246
CHAPTER 29 -- Harvest of Fertilized One-Cell Rat
......Page 250
CHAPTER 30 -- Implantation of Microinjected Eggs
......Page 254
CHAPTER 31 -- Cryopreservation of Transgenic Rat Lines
......Page 258
CHAPTER 32 -- Production of Transgenic Sheep
......Page 260
CHAPTER 33 -- Cryopreservation of Transgenic Sheep Lines
......Page 292
CHAPTER 34 -- Isolation of Genomic DNA from Tail Tissue
......Page 294
CHAPTER 35 -- Slot Blotting of Genomic Tail DNA
......Page 298
CHAPTER 36 -- Genomic Analysis of Transgenic Animals by the Polymerase Chain Reaction
......Page 302
CHAPTER 37 -- Genomic Analysis of Transgenic Animals --
Southern Blotting......Page 308
CHAPTER 38 -- The Preparation of RNA from the Tissues of Transgenic Animals
......Page 313
CHAPTER 39 -- Northern Blotting
......Page 319
CHAPTER 40 -- RNA Slot Blotting
......Page 323
CHAPTER 41 -- Analysis of Gene Expression by PCR
......Page 327
CHAPTER 42 -- Nuclear Run-On Analysis of Transcription
......Page 337
CHAPTER 43 -- S1 Nuclease Protection Mapping
......Page 345
CHAPTER 44-- Primer Extension
......Page 355
CHAPTER 45 -- In Situ Hybridization Analysis of Transgenic Tissues
......Page 361
CHAPTER 46-- Immunohistochemical Analysis of Transgene Expression
......Page 377
CHAPTER 47 -- Postmortem Examination of Transgenic Mice
......Page 389
CHAPTER 48 -- Histological Examination of Transgenic Mice
......Page 395
CHAPTER 49 -- Reporter Enzyme Assays
......Page 401
CHAITER 50 -- Restriction Endonuclease Digestion of DNA
......Page 407
CHAPTER 51 -- Agarose Gel Electrophoresis
......Page 413
CHAPTER 52 -- Working with Radioactivity in the Transgenic Animal Laboratory
......Page 419
CHAPTER 53 -- Random Primed Labeling of DNA
......Page 425
CHAPTER 54 -- Filter Hybridization
......Page 429
CHAPTER 55 -- DNA Transfection
......Page 433