| PART
I |
CONSTRUCTION OF
A RECOMBINANT PLASMID
|
|
| Jan 20 |
Course Introduction
Lab Procedures and Lab Safety
Care and use of the Rainin Pipetman
Problem Set: Common Calculations
in the Molecular Biology Laboratory
Preparation of DNA insert via PCR
|
|
| Jan 27 |
Agarose Gel Electrophoresis of DNA insert
Go over Problem Set
|
|
| Feb 3 |
Perform Cloning Reactions
Transform Competent E. coli cells with Cloning Reactions
|
Preliminary
Project Proposals Due
|
| Feb 10 |
Plasmid minipreps
Assessment of Recombinant Plasmids by Restriction Enzyme Digestion
Restreaking Miniprep Cultures
|
|
| Feb 17 |
Gel Electrophoresis of Restriction Enzyme Digests
Data Analysis
Restreaking Miniprep Cultures
|
|
PART
II |
OVEREXPRESSION
OF RECOMBINANT PROTEIN IN BACTERIAL HOST
|
|
| Feb (23 and) 24 |
Transform Competent E. coli expression host cells with Recombinant Plasmid
Bacterial Cell Culture
Induce Expression of Recombinant Protein
Preparation of Samples for Electrophoresis
|
First Lab Report
Due: Construction of Recombinant Plasmids
|
| March 3 |
|
Final Project Proposals Due
|
| March 10
March 17
|
SPRING BREAK |
|
| March 24 |
SDS Gel Electrophoresis
Western Blotting
|
|
| March (30 and) 31 |
Probing Blots with Anti-His tag Antibody
Development of Western Blot
|
|
| PART
III |
INDEPENDENT RESEARCH
PROJECTS
|
Guidelines
and Deadlines
|
| April 7 |
Begin Exclusive Work on Independent
Projects
|
Second Lab Report Due: Detection of Recombinant Protein Expression By Western Blotting
|
| April 14 |
Independent Projects Continue
|
|
| April 21 |
Independent Projects Continue
|
|
| April 28 |
Independent Projects Continue
|
|
| May 6 |
Oral Presentations on Independent
Projects
|
|
| **May 9 (Monday)** |
|
|