Pulsed field gel electrophoresis a practical guide / Bruce Birren, Eric Lai.

• Front Cover; Pulsed Field Gel Electrophoresis; Copyright Page; Table of Contents; PREFACE; CHAPTER 1. INTRODUCTION TO CONVENTIONAL AND PULSED FIELD GEL ELECTROPHORESIS; 1.1 Conventional Gel Electrophoresis; 1.2 Pulsed Field Gel Electrophoresis; 1.3 Terms That Have Been Used in Pulsed Field Gel Electrophoresis; 1.4 Microscopic Observations of DNA Molecules in Pulsed Field Gel Electrophoresis; 1.5 Pulsed Field Gel Electrophoresis Nomenclature; References; CHAPTER 2. SETTING UP A PULSED FIELD GEL ELECTROPHORESIS SYSTEM; 2.1 Development of Pulsed Field Gel Electrophoresis Apparatuses. 
• 2.2 Description of Components in Pulsed Field Gel Systems2.3 Selecting a Pulsed Field Gel System; 2.4 Building Your Own Pulsed Field Gel System; References; CHAPTER 3. PREPARATION OF DNA FOR PULSED FIELD ANALYSIS; 3.1 Working with Large DNA; 3.2 Size Markers for Pulsed Field Gels; 3.3 Recipes for Solutions and Reagents Used; 3.4 Procedures for Preparation of Pulsed Field Gel Size Markers; 3.5 Procedures for Preparation of Yeast Chromosomes; 3.6 General Considerations for Preparation of Mammalian DNA; 3.7 Procedures for Preparation of Mammalian Cells for DNA Isolation. 
• 3.8 Preparation of DNA from Mammalian Cells3.9 Preparation of High Molecular Weight DNA from Bacteria; 3.10 Preparation of DNA from Mycoplasma; 3.11 Preparation of DNA from Protozoa; 3.12 Preparation of DNA from Caenorhabditis elegans; 3.13 Preparation of DNA from Drosophila; 3.14 Preparation of Neurospora Chromosomes; 3.15 Preparation of DNA from Dictyostelium; 3.16 Preparation of DNA from Plants; References; CHAPTER 4. POURING GELS AND HANDLIN G SAMPLES; 4.1 Introduction; 4.2 Dissolving Agarose; 4.3 Running Buffers; 4.4 Casting Gels; 4.5 Different Types of Agarose; 4.6 Loading Samples. 
• 4.7 Starting a Run4.8 Handling Gels; 4.9 Staining, Visualization, and Photography of Pulsed Field Gels; Reference; CHAPTER 5. RESTRICTION ENZYME DIGESTION OF LARGE DNA FRAGMENT SINAGAROSE; 5.1 Introduction; 5.2 Selection of Restriction Enzyme Buffers; 5.3 Amount of Restriction Enzymes Required for Complete Digestion of Agarose-Embedded DNA; 5.4 General Rules on Restriction Enzyme Selection for Pulsed Field Gel Electrophoresis; 5.5 Procedures for Digesting DNA in Agarose; 5.6 Importance of Negative Controls in Restriction Enzyme Digestions of Large DNA. 
• 5.7 Liquid Digestion Procedure for Generating DNA Fragments Smaller Tha n 500 kb5.8 Solid Plug Digestion Procedure for Generating DNA Fragments Larger Than 500 kb; 5.9 Partial Restriction Enzyme Digestion; 5.10 Partial Digestion by Limiting Reaction Time; 5.11 Partial Digestion by Limiting Restriction Enzyme Concentration; 5.12 Partial Digestion by Limiting Magnesium Concentration; 5.13 Partial Digestion by Methylase Competition; References; CHAPTER 6. SWITCH INTERVALS A N D RESOLUTION IN PULSED FIELD GELS; 6.1 Introduction to Field Switching; 6.2 Longer Switch Intervals for Larger Molecules. 
• 6.3 Switch Intervals and Fragment Resolution. 

• Pulsed Field Gel Electrophoresis: A Practical Guide is the first laboratory manual to describe the theory and practice of this technique. Based on the authors' experience developing pulsed field gel instruments and teaching procedures, this book provides everything a researcher or student needs to know in order to understand and carry out pulsed field gel experiments. Clear, well-tested protocols assume only that users have a basic familiarity with molecular biology. Thorough coverage of useful data, theory, and applications ensures that this book is also a lasting resource for more adv

Ämnesord

Pulsed-field gel electrophoresis  -- Technique -- Handbooks, manuals, etc. (LCSH)

DNA  -- Analysis -- Handbooks, manuals, etc. (LCSH)

Publikations- och innehållstyp

541.372 (DDC)

Uceefa (kssb/8 (machine generated))