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Team:UCSF/Notes/Timeline
From 2010.igem.org
Responsibilities of Team Members
| Member | Lab Work | Other Responsibilities |
|---|---|---|
| Lianna Fung | Cloning, Transfections | Wiki Content |
| Hannah Yan | Cloning | Wiki Content |
| Ethan Chan | Cloning, Cell Culturing, Transfections,Killing Assay | Wiki Content |
| Ryan Liang | Cloning, Cell Culturing, Transfections, Antibody Staining, Killing Assay, Reporter Assay | Wiki Design |
| Crystal Liu | Cloning, Killing Assay | Presentation, Wiki Design |
| Carmen Zhou | Cloning, Transfections, Killing Assay | Wiki Content |
| John Elam | Cloning, Transfections, Killing Assay | Presentation |
| Connor Grant | Cloning | Wiki Content |
| Samuel Zorn | Cloning, Antibody Staining, Reporter Assay | Presentation |
| Eric Wong | Cloning, Cell Culturing, Transfections, Antibody Staining, Killing Assay | Wiki Content |
| Min Lin | Cloning, Transfections, Antibody Staining, Reporter Assay | Wiki Content, Wiki Design, EVERYTHING |
Project Timeline
FEBRUARY
17th – Arrival of NK cell lines – NKL, YTS/eco, KHYG1
18th – Ryan and Ethan begin NK cell cytotoxicity research on cell lines NKL, KHYG-1, NK92MI & YTS/eco
MARCH
16th – Ryan and Ethan starts optimization of NKL cell line
18th – Ryan and Ethan starts optimization of KHYG-1 cell line
APRIL
9th – Troubleshooting of FACS readout due to Propidium Iodide staining
16th – Reduction of Propidium Iodide, cell concentration, and event readout, FACS analysis continues.
20th – Ryan and Ethan starts optimization of NK92MI cell line
22nd – Ryan and Ethan starts optimization of YTS/eco cell line
29th – Second round of optimization begins; NK92MI cell line is discontinued
30th – Preparation of presentation for May 4th meeting with other iGEM students
MAY
4th – Ryan, Ethan and advisor meet with other iGEM students from Abraham Lincoln High School
13th – Ethan and Ryan gain full responsibility of NKL, KHYG-1, YTS/eco & K562 cell lines
18th – Arrival of Anti-Meso/CD19 plasmids from Mike Milone (University of Pennsylvania)
JUNE
11th – Arrival of K562 cell line from Mike Milone (UPenn)
14th – Full local iGEM team arrive at UCSF and begin 2-week boot camp session.
15th – Seminar by Raquel G. on immune response, signaling cascades, feedback loops, receptor adaptors, and general cancer detection
16th – Seminar by Derek W. on cellular cytoskeleton, actin system, microtubules and other related proteins
17th - Seminar by Daniel H. on cell death, killing process, cytotoxic agents and inducers of apoptosis
18th – Seminar by Reid W. on logic gates, combinatorial qualities of proteins and behavioral changes
21st – Seminar by David P. on modularity, synthetic biology, and Boolean gates
22rd – iGEM Team challenge; brainstorming and consulting with advisors and grad students
24th – General project in mind: granzyme linking, stronger signaling and greater arsenal
25th – Lab safety training begins, first set of primers are designed, first set of source plasmids are ordered, and lab protocols are learned.
28th – Lab work begins
JULY
1st – International student, Min L. arrives from China. Source plasmids arrive for transformation
6th – Primers were incorrect so all labwork had to be redone and primers had to be reordered
8th – Transformations of first bulk source plasmids begin
9th – Tilden Park BBQ with Berkeley
12th – Gel Extraction, Restriction Digests & Colony PCR begin
13th – Minipreps begin
14th – Sequencing of parts begin
16th – Sequences show contamination in CD28, Grb2, and mDAP10 plasmids, team project description due
19th – Ly49 mRNA Contamination, restriction digest failures, and mix ups become prevalent
22nd – Second source plasmid bulk arrives
23rd – Positively sequenced primers are preserved in glycerol.
24th – Ryan, Ethan, and Eric starts first dry run transfection in NKL cell line with anti-Mesothelin CAR for killing assays
26th – AB parts were incorrect and Sam and Connor redo these parts
28th – iCLEM visit
29th – Carmen and Ryan makes BD backbone and retrieve AB-start codon (UCSF iGEM 2009) for ligations
30th – Ryan, Ethan, and Eric starts transfection in NKL cell line with anti-CD19 for killing assays
AUGUST
2nd – Ligations of AB, BC, CD parts
3rd – Granule project theories with advisors
6th – Ryan and Min start antibody staining and blocking for killing assay
7th – Killing assay used the wrong plasmids, redo with correct plasmid
9th – Transfection Assay Optimization continues
16th – Ryan, Ethan, Sam and Eric leave for school
17th – Sam and Min begins oligo synthesis
19th – Ethan and Eric continue killing assay with correct plasmid
22nd – Hannah leaves for school
23rd – Granule oligos synthesized
27th – Begin endo-free maxipreps of completed constructs
30th – Start pH sensitive GFP, LIMP, LAMP, and eGFP for granule side project
SEPTEMBER
5th – Connor leaves for UCSD for soccer tryouts
6th – pH sensitive GFP, LIMP, LAMP granule project discontinued
9th – Killing assay discontinued, transfection efficiency too low
20th – Connor, Crystal, John, Carmen, and Lianna leave for school
21st – T-cell activation assays begin, presentation draft created
26th – Connor returns from UCSD
27th – Min starts first granule project assay on eGFP
OCTOBER
3rd – Min leaves for China
4th – Sam takes over granule project and begins cell imaging
7th – End of construct production, 59 constructs completed from endo-free maxipreps, 17 have been preserved in glycerol
8th – Ryan starts imaging on live KHYG1 for images of killing; results for T-cell activation assays data is revealed, final T-cell activation assay
12th – Preparation for presentation begins
15th – Killing imaging completed
20th – Granule project: eGFP imaging completed
23th – NorCal Jamboree with Stanford, UC Davis, and UC Berkeley iGEM teams
27th – iGEM Wiki freeze
30th – Continue presentation preparation
NOVEMBER
5th – iGEM 2010 Jamboree!
