Molecular Biology / Wetlab

September 11-13 2018

Module leaders:
Paul Thomas (Frost Lab)
Jason Town (Weiner Lab)

TAs:
Lakshmi Miller-Vedam (Frost/Weissman Lab)
Kyle Lopez (Southworth Lab)

Description:
In this 2.5-day hands-on lab module we will use basic biochemistry techniques to isolate and purify multiple fluorescent proteins. We will used these purified fluorescent proteins to explore protein folding and unfolding with various chemical and  thermal denaturants. Students will compare these proteins stability and refolding capacity and present their findings in a quick, but formal talk.

In addition, students will prepare 10min chalk talks (5 min questions) on relevant protein folding and biochemistry papers which will be provided ahead of time.


Paper Groups:


Group 3: Garrett & Hayarpi & Matthew: Protein Folding and Chaperones
Group 4: Jack & Laura & Nicholas: Protein Expression and Localization
Group 5: Daniel & Stephanie & Maurisa: Unfolded Protein Reponse

Group 6: Miriam & Laurel & Bryan: Cell Division

Tentative Lab Groups:
Group 1: Christina & Elizabeth & Maria
Group 2: Calla & Elissa & Daniel
Group 3: Stephanie & Garrett & Nicholas
Group 4: Maurisa & Laurel & Ajikarunia
Group 5: Matthew & Miriam & Jack
Group 6: Bryan & Laura & Hayarpi



Schedule:

September 11th, 2018

Time

Activity

9:00 - 9:30

Coffee+Thaw Lysates
9:30 - 10:00 Balance Centrifuge Tubes
10:00 - 11:00 Spin Down Lysates, Equilibrate Beads
11:30 - 12:30 Bind Protein to Beads
LUNCH
12:30 - 13:30 Wash and Elute Protein
13:30-14:45 Run Gels of Purification
14:45-end of day Quantify Protein Yield
Prepare Dialysis for Next Day
Talk with your paper group for tomorrow

September 12th, 2018

Time

Activity

9:00 - 9:30

Coffee
9:30 - 11:00 Denature your protein
11:00-12:30 10min Chalk Talks on papers, 5 min questions
12:30-13:30 LUNCH
13:30-end of day Assay Fluoresence after denaturation/refolding
Analyze data for tomorrow

September 13th, 2018

Time

Activity

9:00 - 10:30

Prepare Presentations
(12 min slideshow, 3 min questions)
10:30-12:00 Present
12:00-13:00LUNCH