Difference between revisions of "Team:William and Mary/Notebook"
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Revision as of 21:06, 17 October 2018
Notebook
Week 1: 180528 -180604
- Began cloning 3G parts based off the library sent from the Murray lab
- Researched literature on dual inducible promoters
- Sent out several cloned 3G parts for sequencing
- Planned out several IFFLs to test.
Week 2: 180604 -180610
- Continued cloning 3G parts onto our standard WM Pad backbone
- Redesigned primers for cloning 3G promoter parts, as previous ones were faulty
- Received results from sequencing, confirmed parts
- Beginning to assemble circuits using 3G assembly
- Making inducible reporter construct and an mf-Lon based IFFL
Week 3: 180611 -180617
- Completed cloning the portion of our comprehensive 3G assembly part library needed for our circuits
- Adding remaining 3G parts from Murray’s lab to our library
- Working on designing and building a dual inducible promoter and low copy backbone for assembly
- Confirmed minipreps based on sequencing results
- Started plate reader for experiments on circuits with IPTG and ATC inducer
- Started using FACS machine for experiments
Week 4: 180618 - 180624
- Microscopy training with Jacob Sawyer, advanced imaging specialist from Nikon
- Spin down inducer tests to see if we can remove inducer from media to create pulses
- Plate reader experiment of pulse timing in different conditions
- FACs experiment of how long cells can be incubated on ice without changing results
- Decided to stop sequencing 3G parts and start functionally screening them instead
- Set up our first Golden Gate reaction
- Madilyn & Friends Outreach event
Week 5: 180625 -180701
- Continue designing and building circuits
- Continue inducer spin down tests for plate reader experiment
- Reached out to St. Andrews to solidify collaboration partnership
- Considering alternatives for the inducer spin down tests due to multiple complications
- Looked into light-induced gene expression systems
- SEP 8th graders
Week 6: 180702 -180708
- Experienced some leakiness with our FACS machine, spent time fixing this
- Abandoned light-induced gene expression system, as it appears impractical in lab and less relevant to in vivo systems
- Began looking into temperature-induced circuits
- Continue inducer spin down tests
- Making different variants of the same circuit with different strength repressors
- Made and characterized Oscillator Circuit
- Cell sorting with GS, image dissociated cells on the slides
- Started Interlab Study measurements
Week 7: 180709 -180715
- Successfully cloned our entire 3G library, except recently added parts
- Demonstrated cells can be frozen in 15% glycerol without changing results
- Successfully tested whether inducers can be removed from media
- Using parts from last year, tested an IFFL with a single 20 minute pulse
- Tested an IFFL with a single 40 minute pulse
- Began assembling and testing circuits utilizing temperature as an inducer (TS-C1 system)
- Attended the Mid-Atlantic Meetup hosted by University of Maryland
- Submitted Interlab measurements
Week 8: 180716 -180722
- Reached out to UVA for collaboration
- Continued with plate reader experiments
- Continued testing heat inducible circuits
- Cloned and transformed Oscillator parts for functional testing
- Medical Explorers Outreach Event Visit Day 1 and 2
- Building with Biology event
- Camp Launch
Week 9: 180723 -180729
- Contacted University of Pittsburgh for collaboration
- Continued plate reader experiments
- Testing lower temperature inductions
- Successfully created working high copy inducible parts and possible working low copy parts
- Addressing issues with microscopy
- Plus S Ice Cream and iGEM
Week 10: 180730 -180805
- Continuing plate reader experiments
- Interlab data got rejected due to accidentally switching the Od600 and fluorescence in the form. Minor issue, easily resolved and submitted
- Investigated parameters for oscillator
- Sent collaboration package to UVA and University of Pittsburgh
- Started considering clontegration and requisite parts and primers
- First MUN Meeting
Week 11: 180827 -180902
- Designed primers for pOSIP 3G amplification
- Continued plate reader experiments with heat inducible circuits
- Updated safety form
Week 12: 180903 -180909
- Continued plate reader experiments
- Focused on testing variations of Tlp- heat sensitive promoter
- Collaboration data was received from UVA and University of Pittsburgh
- Design logo
Week 13: 180910 -180916
- CTested circuit with two different promoters, tlp mflon and tlp mscarlet
- Hurricane Florence hit, school was evacuated and closed from Tuesday to Sunday
- No wetlab was able to commence
- Analyzed data from integrated circuits
- Presentation at Windsor Mead Nursing Home
Week 14: 180917 - 180923
- Picked up after Hurricane Florence
- Made competent flped cells
- Tested circuits transformed in pre-flped and flped cells
- Tested new ptd tag
- Experiments with integrated cells
- Created parts pages for UVA parts
- Presentation at Williamsburg Landing Nursing Home
- Poster Presentation at the Women’s Weekend
Week 15: 180924 - 180930
- Plate reader experiment with older parts- tsc1 mscarlet with tlp mflon
- Tested more ptd tag parts
- Continue experiments with integrated cells
- Analyzed different strength 3k3 parts
- Outbreak presentation with guest entrepreneurship speaker
Week 16: 181001 - 181007
- Plate reader experiment of combined circuits with tlp mflon and tlp mscarlet
- Cloned multiple circuits to be sequenced and submitted
- Presentation to Model UN
Week 17: 181008-181017
- Wrapped up on outstanding wetlab tasks
- Worked on Wiki!
- Ladies in the Lab