Team:William and Mary/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