Difference between revisions of "20.109(S08):Module 2"
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[[20.109(S08):Start-up protein engineering | Module 2 Day 1: Start-up protein engineering]]<br> | [[20.109(S08):Start-up protein engineering | Module 2 Day 1: Start-up protein engineering]]<br> | ||
| − | + | Students dissect pericam sequence into constituent parts. | |
| − | + | Read about CaM-M13 and use protein viewer to choose targets, then plan primers for SDM. | |
| − | + | Also get calcium titration curve for WT plasmid using Nanodrop? | |
| − | + | [[20.109(S08):Site-directed mutagenesis| Module 2 Day 2: Site-directed mutagenesis]]<br> | |
| − | + | ||
| − | [[20.109(S08):Site-directed mutagenesis| Module 2 Day | + | |
(Staff or students do transformation/miniprep/transformation work? Timing issues, may take another lab day...) | (Staff or students do transformation/miniprep/transformation work? Timing issues, may take another lab day...) | ||
| − | [[20.109(S08):Characterize protein expression| Module 2 Day 4: Characterize protein expression]]<br> | + | [[20.109(S08):Characterize protein expression| Module 2 Day 3: Induce protein expression]]<br> |
| + | |||
| + | (IPTG induction - meanwhile, discuss journal article; collect and pellet samples) | ||
| + | |||
| + | [[20.109(S08):Protein binding assays| Module 2 Day 4: Characterize protein expression]]<br> | ||
| − | + | Extract protein and SDS-PAGE; purify protein and quantify amount. | |
| − | + | Note: week off between day 4 and day 5 of lab. | |
| − | ( | + | [[20.109(S08):Sequence analysis| Module 2 Day 5: Protein binding assay and Sequence analysis]]<br> |
| − | + | Calcium titration curve, using plate reader. Then come back and start data analysis. | |
| − | ( | + | [[20.109(S08):Name| Module 2 Day 6: Name]]<br> |
| − | + | MATLAB modeling day? Other reading or work with calcium indicators? | |
[[20.109(S08):Name| Module 2 Day 7: Name]]<br> | [[20.109(S08):Name| Module 2 Day 7: Name]]<br> | ||
| − | ( | + | Options as Day 6. Could have two days of intro/design, but this means pellets are sitting around for a week before protein extraction (better or worse than protein sitting around for a week?) |
[[20.109(S08):Student presentations| Module 2 Day 8: Student presentations]]<br> | [[20.109(S08):Student presentations| Module 2 Day 8: Student presentations]]<br> | ||
Revision as of 20:40, 20 November 2007
Module 2 Day 1: Start-up protein engineering
Students dissect pericam sequence into constituent parts.
Read about CaM-M13 and use protein viewer to choose targets, then plan primers for SDM.
Also get calcium titration curve for WT plasmid using Nanodrop?
Module 2 Day 2: Site-directed mutagenesis
(Staff or students do transformation/miniprep/transformation work? Timing issues, may take another lab day...)
Module 2 Day 3: Induce protein expression
(IPTG induction - meanwhile, discuss journal article; collect and pellet samples)
Module 2 Day 4: Characterize protein expression
Extract protein and SDS-PAGE; purify protein and quantify amount.
Note: week off between day 4 and day 5 of lab.
Module 2 Day 5: Protein binding assay and Sequence analysis
Calcium titration curve, using plate reader. Then come back and start data analysis.
MATLAB modeling day? Other reading or work with calcium indicators?
Options as Day 6. Could have two days of intro/design, but this means pellets are sitting around for a week before protein extraction (better or worse than protein sitting around for a week?)
Module 2 Day 8: Student presentations