LABORATORY 6. MOLECULAR BIOLOGY
OVERVIEW
In this laboratory, you will investigate some basic principles of genetic engineering.
Plasmids containing specific fragments of foreign DNA will be used to transform Escherichia
coli cells, conferring antibiotic (ampicillin) resistance. Restriction enzyme digests
of phage lambda DNA also will be used to demonstrate techniques for separating and
identifying DNA fragments using gel electrophoresis.
OBJECTIVES
Section A: Before doing this laboratory you should understand:
- how gel electrophoresis separates DNA molecules present in a mixture
- the principles of bacterial transformation
- the conditions under which cells can be transformed
- the process of competent cell preparation
- how a plasmid can be engineered to include a piece of foreign DNA
- how plasmid vectors are used to transfer genes
- how antibiotic resistance is transferred between cells
- how restriction endonucleases function
- the importance of restriction enzymes to genetic engineering experiments
Section B: After doing this laboratory you should be able to:
- use plasmids as vectors to transform bacteria with a gene for antibiotic resistance in a
controlled experiment
- demonstrate how restrictions enzymes are used in genetic engineering
- use electrophoresis to separate DNA fragments
- describe the biological process of transformation in bacteria
- calculate transformation efficiency
- be able to use multiple experimental controls
- design a procedure to select positively for antibiotic resistant transformed cells
- determine unknown DNA fragment sizes when given DNA fragments of known size
LAB GUIDE
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Lab 6- Electrophoresis
