The Virtual Lab Book by Dr. Stephanie Dellis College of Charleston

 

Introduction

    Welcome to the Virtual Lab Book!

The Virtual Lab Book was prepared as a guide for students in the Molecular Biology Lab at the College of Charleston. However, all students are welcome to use it.

Some sections of the Lab Book are image-intensive, and may be frustrating to use with slower modems. I have tried to keep the images at each link to a minimum. However one result of this is more links. The Virtual Lab Book images are meant as a visual on-line resource and are not really designed for printing out. The "readings" however are meant to be printed.

The Adobe pdf files require the free Acrobat reader.

Contents

1. Safety in the Molecular Biology Lab (pdf)

2.  Basic Skills in Molecular Biology (pdf)

Topics:

• Micropipettors and Pipetting Skills
• Using Bacteria in Molecular Biology

Protocol: BasicSkills (pdf)

Web pages:

• Plasmid antibiotic resistance
• How to streak a plate
• Streak plate examples:  "the good, the bad, and the ugly"

3.  Restriction Digestion and Agarose Gel Electrophoresis (pdf)

Topics:

• DNA Cloning
• Restriction Enzymes and Their Use
• Plasmids
• Cloning Scheme
• Separating by Electrophoresis
• Ligation of a Gene into a Vector

Protocol: RE_Gels (pdf)

Web pages:

• Loading an Agarose Gel

4. Competent Cells and Transformation (pdf)

Topics:

• Bacterial competence
• Identification of positive clones

Protocol: CompCells_Transf (pdf)

Protocol: Starting Overnight Cultures

Web pages:

• How to Spread a Plate
• Spreading Examples

5. Minipreparation of Plasmid DNA (pdf)

Topics:

• Minipreparation of plasmid DNA
• Restriction digestion to identify positive clones

Protocol: Miniprep (pdf)

Web page:

• Controls in Molecular Biology

6. PCR and Thermalcycling (pdf)

Topics:

• Isolation of genomic DNA
• Thermalcycling
• Polymerase chain reaction

Protocol 1: DNA Isolation (pdf)

Protocol 2: PCR (pdf)

7. 16s rDNA Sequencing to Identify Unknown Microorganisms (pdf)

Protocol: rDNA isolation (pdf)

8. Cycle Sequencing (pdf)

Topics:

• Cycle sequencing
• Flourescence detection

9. Interpreting DNA Sequence (pdf)

10. Constructing a Phylogenetic Tree (pdf)

11. Human DNA Typing (handout)

Protocol: Isolation of cheek cell DNA (pdf)

12.  Polyacrylamide Gel Electrophoresis

Web pages:

• Preparing the Acrylamide Gel
• Assembling the Gel Apparatus