Beginning Molecular Biology Laboratory Manual

CHAPTER 1: General Laboratory Methods

Safety Procedures
Preparation of Solutions
Disposal of Buffers and Chemicals
Equipment Micropipets

Using a pH meter
Autoclave operating procedures
Operating instructions for spectrophotometer

Working with DNA
Sterile Technique
CHAPTER 2: Instructions for Notebook Keeping

CHAPTER 3: Computer User’s Guide

UNIX commands
File Transfer
The World Wide Web
The Wisconsin Package – gcg.

CHAPTER 4: Molecular Biology Methods
M.1: Preparation of genomic DNA from bacteria
M.2: PCR amplification of DNA
M.3: Restriction enzyme digestion of DNA
M.4: Phenol/chloroform extraction of DNA
M.5: Ethanol precipitation of DNA
M.6: Agarose gel electrophoresis
M.7: Transformation of E. coli by electroporation

M.8: Wizard PCR preps DNA purification system
M.9: Alternate method for purifying DNA from agarose gels
M.10: Transfection of mammalian cells using Lipofectamine (LTI)
M.11: Southern blotting
M.12: RT-PCR Protocol
M.13: Preparation of sequencing gels
M.14: Isolation of RNA from mammalian cells using RNAZOL (Teltest)

CHAPTER 5: Tissue Culture Methods
Types of cells grown in culture
Work area and equipment
Preservation and storage
Maintenance
Safety considerations
Tissue culture methods
Determining cell counts