20.109(S08):Module 2: Difference between revisions
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[[20.109(S08):Site-directed mutagenesis| Module 2 Day 2: Site-directed mutagenesis]]<br> | [[20.109(S08):Site-directed mutagenesis| Module 2 Day 2: Site-directed mutagenesis]]<br> | ||
(Staff or students do transformation/miniprep/transformation work? Timing issues, may take | (Staff or students do transformation/miniprep/transformation work? Timing issues, may take a second lab day...) | ||
[[20.109(S08):Characterize protein expression| Module 2 Day 3: Induce protein expression]]<br> | [[20.109(S08):Characterize protein expression| Module 2 Day 3: Induce protein expression]]<br> | ||
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[[20.109(S08):Protein binding assays| Module 2 Day 4: Characterize protein expression]]<br> | [[20.109(S08):Protein binding assays| Module 2 Day 4: Characterize protein expression]]<br> | ||
Extract protein and SDS-PAGE; purify protein and quantify amount. | Extract protein and perform SDS-PAGE; purify protein and quantify amount. | ||
Note: week off between day 4 and day 5 of lab. | Note: week off between day 4 and day 5 of lab. | ||
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Calcium titration curve, using plate reader. Then come back and start data analysis. | Calcium titration curve, using plate reader. Then come back and start data analysis. | ||
Also sequence analysis. | |||
[[20.109(S08):Name| Module 2 Day 6: Name]]<br> | [[20.109(S08):Name| Module 2 Day 6: Name]]<br> | ||
Revision as of 13:42, 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 a second 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 perform 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.
Also sequence 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?)
