Started preparing Ampicillin stocks:

Materials:
Ampicillin(from bio lab)
MilliQ(nuclease free water) update: used sterile water
Falcon tubes
250 mL bottle
Microcentrifuge tubes

Started sponsorship emails: refer to sponsorship spreadsheet for more information

Continued research on project ideas:
Anaphalactic shock idea
TTSS to cure cancer idea

Advisor Meeting 3-4pm:

The advisors suggested not developing ideas much but instead focusing on brainstorming and a more scattershot approach where many ideas are discussed but not developed as much

Deemed responsibilities:
Yuan: ordering, budget, wiki
Brian: wiki, human practices
Tae: autoclaving
Grant: sponsorship, notebook
Mike: human practices, twitter
Sarah: autoclaving, human practices
Lajja: sponsorship, autoclaving

Sarah and Tae created the CCMB buffer solution(protocol)

Brian and Yuan obtained top10 seed stock cells from Leonard’s lab. They dropped the tip of cells into overnight tubes, filled with 5 mL of SOB, and incubated and shook at 200 rpm at 37 degrees celsius overnight.

The morning was spent continuing research on new project ideas. As of now, there are no well defined project ideas.

Brian placed 1 mL of incubated bacteria into 100 mL SOB in erlenmeyer flask, placed the Erlenmeyer flaskin the shaker at 200rpm at 37 degrees celsius and continually checked the OD of the cells.

Protocol taken from “Molecular Cloning” Sambrook and Russell: modified MgCl2-CaCl2 solution to CCMB buffer solution

2:00 pm cells placed in shaker
4:10 pm OD .189
5:45 pm OD .745

The OD of the cells was much too high and the cells were discarded. An OD of .400 needs to be reached next time.

TContinued research regarding 3 main projects:
Methanotrophs turning methane into methanol. Contacting professor at northwestern with significant experience with methanotrophs
3d DNA scaffolding. Professor Leonard suggested looking into DNA and RNA scaffolding.
Bacterial signaling chain of luciferase and halorhodopsin.

Yuan made SOB broth following molecular and iGEM protocol
100 mL SOB
.5 mL MgCl2
autoclave 20 minutes

Continued making Top10 competent cells after yesterday’s mishap. Today we will monitor the OD of the cells much more closely.
12:45 pm OD .100
1:06 pm OD .151
1:25 OD .221
1:45 OD .325
1:50 OD .348

10 minutes in centrifuge tube on ice
10 minutes centrifuging @ 4 degrees celsius 4000 rpm
Resuspended bacteria in CCMB buffers 30mL by electric pipet after decanting SOB
10 minutes centrifuging same as above
Resuspended in 2mL CCMB by lightly shaking the tube
Placed in 4 1mL aliquots in -80 freezer

Continued research regarding the three project ideas (methanotrophs, dna scaffolding, bacterial signaling chain)

Continued making Top10 competent cells

Yuan and Brian started developing the wiki page

Mike reactivated last year’s NUiGEM twitter account and is now the social media manager