20.109(S08):Module 2

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.

Module 2 Day 6: Name

MATLAB modeling day? Other reading or work with calcium indicators?

Module 2 Day 7: Name

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