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!