| Week |
Monday |
Tuesday |
Friday |
Laboratory |
| 1 |
Feb 2 |
Feb 3 |
Jan 30 |
Introduction and enzyme assays |
| 2 |
Feb 9 |
Feb 10 |
Feb 6 |
Discussion of purification methods.
Decision on which methods and variations to be used by each team.
Make buffers and equilibrate gels. |
| 3 |
Feb 20 (Fri) |
Feb 17 |
Feb 13 |
Purification begins: Filter egg-white
and perform ion-exchange chromatography. |
| 4 |
Feb 23 |
Feb 24 |
Feb 27 |
Purification begins: Collect fractions
and enzyme assay of fractions. |
| 5 |
Mar 1 |
Mar 2 |
Mar 5 |
Protein enzyme assays of fractions
and SDS-PAGE |
| 6 |
Mar 8 |
Mar 9 |
Mar 19 |
Photographing gels and discussion
of purification results |
| 7 |
Mar 12 |
Mar 16 |
Apr 2 |
Project on protein
methods: Rotation I |
| 8 |
Mar 26 |
Mar 30 |
Apr 9 |
Project on protein methods: Rotation
II |
| 9 |
Apr 5 |
Apr 6 |
Apr 16 |
Independent project: Variation
on a protein method |
| 10 |
Apr 12 |
Apr 13 |
Apr 23 |
Independent project continues
|
| 11 |
Apr 26 |
Apr 27 |
Apr 30 |
Write short paper.
|
| 12 |
May 3 |
May 4 |
May 7 |
Presentation of project results.
|
| Week |
Assignment |
| Week 1 |
Bring lab manual and lab notebook
to lab. |
| Week 2 |
Calculate the specific activities
of the lysozyme standard. Read the textbook section on lysozyme. |
| Week 3 |
Outline the approach you will
take to purify lysozyme using a flow diagram. For a batch method,
indicate if lysozyme will reside in the supernatant or precipitate.
For column separations, indicate the likely contents of the run-off
and the subsequent eluted volumens. |
| Week 4 |
Review the progress made during
the first week of purification and decide on your approach for this
week (e.g., method of column elution). |
| Week 5 |
Prepare a dilution scheme for
BSA, the 2 mg/mL standard for the protein assay. Decide on the dilutions
of your unknown for the protein assay. Attempt three different dilutions
to improve chances that the result will fall on the standard curve.
For the enzyme assay, decide on dilutions of your unknowns that contained
protein. |
| Week 6 |
Graph the standard curve for the
protein assay and complete the calculations for all assays. Complete
a data summary table of your results. Be ready to report to the class
the specific activity, the total activity, the % yield, and the fold
purification of your fractions. |
| Week 7 |
Read the sections in the manual
on your assigned protein method and also read the listed references.
NOTEBOOKS ARE DUE. |
| Week 8 |
Read the section in the manual
on your assigned protein method and also read the listed references.
Schedule a meeting with your instructor three days after lab to discuss
independent project and bring a materials list. |
| Week 9 |
Complete a pre-lab plan in your
notebook for your independent project (e.g., calculate how to prepare
appropriate volumes of buffers, reagents, enzymes, etc.) |
| Week 10 |
Complete a pre-lab plan in your
notebook for this second week of lab work. |
| Week 11 |
Schedule a 20-min meeting with
your instructor to discuss your results. Use the remaining lab time
to write your scientific paper. |
| Week 12 |
Presentation of your independent
project. Bring a copy of your draft to class for peer review. |
| Reading Period |
FINAL
PAPER DUE. Also turn in the peer review forms completed
by other students and your first draft. |
