Molecular Biology Lab Manual

 

Julie B. Wolf

Department of Biological Sciences, UMBC

 

 

 

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