CONSTRUCTION OF PLANT BACTERIAL ARTIFICIAL CHROMOSOME (BAC) LIBRARIES: AN ILLUSTRATED GUIDE

TABLE OF CONTENTS

ABSTRACT


INTRODUCTION

CHAPTER 1 Overview

CHAPTER 2 Supplies, equipment, & reagents


VECTOR PREPARATION

CHAPTER 3 Vector isolation

CHAPTER 4 Test ligation

CHAPTER 5 Test transformation

CHAPTER 6 Miniprep & NotI digest


PREPARATION OF INSERT DNA

CHAPTER 7 Isolation of high molecular weight nuclear DNA

CHAPTER 8 DNA analysis

CHAPTER 9 Test restriction digest

CHAPTER 10 Restriction digest

CHAPTER 11 First size selection

CHAPTER 12 Second size selection

CHAPTER 13 Isolation of size-selected DNA from agarose


LIBRARY CONSTRUCTION

CHAPTER 14 Ligation, test transformation, & NotI digest

CHAPTER 15 Mass Transformation

CHAPTER 16 Picking clones

CHAPTER 17 Library replication & storage


APPENDICES

APPENDIX A Using the Gibco BRL Cell-Porator System

APPENDIX B Using the Bio-Rad Model 422 Electroelution System

APPENDIX C Filling 384-well plates using the Genetix QFill2

APPENDIX D The Genetix QBot

APPENDIX E Picking clones using the Genetix QBot

APPENDIX F Library replication using the Genetix QBot


GLOSSARY


ACKNOWLEDGEMENTS


REFERENCES