Methods in Enzymology

Table of Contents

Contributors to Volume 225 xi

Preface xvii

Volumes in Series xix

Hints on Using the Guide to Techniques in Mouse Developentxxxiii

Section I. General Methodology

1. Recordkeeping and Database Analysis of Breeding Colonies 
   Lee M. Silver 3
2. Anethesia and Perioperative Care 
   P.A. Flecknell 16

Section II. Germ Cells and Embryos

1. Culture and Manipulation of Primordial Germ Cells
   Julie E. Cooke, Isabelle Godin, Charles French-Constant, Janet Heasman, and Christopher C Wylie 37
2. Insolation and Culture of Primordial Germ Cells
   Mia Buehr and Anne McLaren 58
3. Isolation and Culture of Oocytes
   John J. Eppig and Evelyn E. Telfer 77
4. Purification, Culture, and Fractionation of Spermatogenic Cells
   Anthony R. Bellvé 84
5. Recovery, Capacitation, Acrosome Reaction, and Fractionation of Sperm
   Anthony R. Bellvé, Wenxin Zheng, and Yordanka S. Martinova 113
6. Strategies and Methods for Evaluating Acrosome Reaction
   Richard A. Cardullo and Harvey M. Florman 136
7. Culture of preimplantation Embryos
   Joel A. Lawitts and John D. Biggers 153
8. Isolation and Culture of Whole Postimplantation Embryos and Germ Layer Derivatives
   Karin Sturm and Patrick P.L. Tam 164
9. Histogenetic Potency of Embryonic Tissues in Ectopic Sites
   Patrick P.L. Tam 190

Section III. Fertilzation

1. Micromanipulation of Gametes and Embryos 
   Jon W. Gordon 207
2. In Vitro Capacitation and Fertilization
   Lynn R. Fraser 239
3. In Vitro Fertilization
   Jeffrey D. Bleil 253

Section IV. Gene Expression: Messenger RNA

1. Radioisotope Labeling of UTP and ATP Pools and Quantitative Analysis of RNA Synthesis
   Kerry B. Clegg and Lajos Pikó 283
2. Quantitative Analysis of Specific Messenger RNAs by Hybridization Using RNA Probes
   Kent D. Taylor and Lajos Pikó 283
3. Quantitative Analysis of Specific Messenger RNAs by Ribonuclease Protection
   Ross A. Kinloch, Richard J. Roller, and Paul M. Wassarman 294
4. Analysis of Messenger RNA
   Mayi Y. Arcellana-Panlilio and Gilbert A. Schultz 303
5. Quantitative Analysis of Specific Messenger RNAs in Small Numbers of Preimplantation Embryos
   Michelle F. Gaudette, Qiu Ping Cao, and William R. Crain 328
6. Quantitative Analysis of Messenger RNA Levels: Reverse Transciption-Polymerase Chain Reaction Single Nucleotide Primer Extension Assay
   J. Singer-Sam and A.D. Riggs 344
7. Microinjecting Antisense Sequences into Oocytes
   Fernando J. Sallés, William G. Richards, Joaquin Huarte, Jean-Dominique Vassalli, and Sidney Stickland 351
8. Detection of Messenger RNA by in Situ Hybridizzation to Tissue Sections and Whole Mounts
   David G. Wilkinson and M. Angela Nieto 361
9. Detection of Messenger RNA byin Situ Hybridization to Postimplantation Embryo Whole Mounts
   Ronald A. Conlon and Bernhard G. Herrmann 373
10. Detection of Messenger RNA by in Situ Hybridization
    David Sassoon and Nadia Rosenthal 384
11. Prepation of Injection Pipettes
    Miriam Miranda and Melvin L. DePamphilis 407
12. Application of Firefly Luciferase to Preimplantation Development
    Miriam Miranda, Sadhan Majumder, Maria Wiekowski, and Melvin L. DePamphilis 412
13. Application of LacZ Gene Fusions to Preimplantation Development
    Murial Vernet, Claire Bonnerot, Pascale Briand, and Jean François Nicolas 434
14. Application of LacZ Gene Fusions to Postimplantation Development
    Claire Bonnerot and Jean-François Nicolas 451

Section VI. Gene Expression: Proteins

1. Two-Dimensional Gel Analysis of Protein Synthesis
   Keith E. Latham, James I. Garrels, and Davor Solter 473
2. One Dimensional Gel Analysis of Histone Synthesis
   Maria Wiekowski and Melvin L. DePamphilis 489
3. Whole-Mount Immunohistochemistry
   Claytus A. Davis 502
4. Techniques for Localization of Specific Molecules in Oocytes and Embryos
   Calvin Simerly and Gerald Schattien 516

Section VII. Gene Expression: Methylation

1. Polymerase Chain Reaction for the Detection of Methylation of a Specific CpG Site in the Gópod Gene of Mouse Embryos 
   Maurizio Zuccotti, Mark Grant, and Marilyn Monk 557
2. Analysis of Methylation and Chromatin Structure
   G.P. Pfeifer, J. Singer-Sam, and A.D. Riggs 567

Section VIII. Gene Identification

1. Construction of Primary and Subtracted cDNA Libraries from Early Embryos
   Jay L. Rothstein, Dabney Johnson, Joel Jessee, Jacek Skowronski, Julie A. DeLoia, Davor Solter, and Barbara B. Knowles 587
2. Consturction of cDNA Libraries from Single Cells
   Gerard Brady and Norman N. Iscove 611
3. Construction and Characterization of Yeast Artifical Chromosome Libraries from the Mouse Genome
   Zoia Larin, Anthony P. Monaco, Sebastian Meier-Ewert, and Hans Lehrach 623
4. Genome Developmentally Regulated Gene Families
   Brian J. Gavin and Andrew P. McMahon 653
5. Cloning Developmentally Regulated Gene Families
   Brian J. Gavin and Andrew P. McMahon 653
6. Screening for Novel Pattern Formation Genes Using Gene Trap Approaches
   David P. Hill and Wolfgang Wurst 664
7. Insertional Mutagenesis by Retroviruses and Promotoer Traps in Embryonic Stem Cells
   Glenn Friedrich and Philippe Soriana 681
8. Two-Dimensional Gel Analysis of Repetitve DNA Families
   Rachel D. Sheppard and Lee M. Silver 701

Section IX. Nuclear Transplantation

1. Transplantation of Nuclei to Oocytes and Embryos
   Keith E. Latham and Davor Solter 719
2. Manipulations of Genetic Constitution by Nuclear Transplantation
   Sheila C. Barton and M. Azim Surani 732

Section X. Transgenic Animals: Pronuclear Injection

1. Production of Transgenic Mice
   Jon W. Gordon 747
2. Factors Influencing Production Frequencey of Transgenic Mice
   Jeffrey R. Mann and Andre P. McMahon 771
3. Surgical Techniques in Production of Transgenic Mice
   Jeffrey R. Mann 782
4. Identification of Transgenic Mice
   Maureen Gendron-Maguire and Thomas Gridley 794

Section XI. Transgenic Animals: Embryonic Stem Cells and Gene Targeting

1. Derivation of Embryonic Stem Cell Lines
   Susan J. Abbondanzo, Inder Gadi, and Colin L. Stewart 803
2. Production of Chimeras between Embryonic Stem Cells adn Embryos
   Colin L. Stewart 823
3. Gene Targeting in Embryonic Stem Cells
   Ramiro Ramírez-Solis, Ann C. Davis, and Allan Bradley 855
4. Simple Scerening Procedure to Detect Gene Targeting Events in Embryonic Stem Cells
   Frank Kõntogen and Colin L. Stewart 878
5. Manipulation of Transgenes by Site-Specific Recombination: Use of Cre Recombinase
   Brian Sauer 890
6. Embryonic Stem Cell Differentiation in Vitro
   Micheal V. Wiles 900
7. Production of Tetraploid Embryos by Electrofusion
   K.J. McLaughlin 919

Section XII. Lineage Analysis

1. Lineage Analysis Using Retroviral Vectors
   Constance L. Cepko, Elizabeth F. Ryder, Christopher P. Austin, Christopher Walsh, and Donna M. Fekete 933

Author Index 961

Subject Index 991