Team:Penn/Notebook
From 2012.igem.org
June 2012 Notebook
Week 1
June 6th
- Set up some lab equipment
- Autoclaved for a while
- Organized biobrick stuff
- Called Vinoo about DNA planning
June 7th
- Transformed Cph8, pLsr, and LuxS
- Placed order with Vinoo
- Developed idea using PGY/PCN system to activate a gene
Week 2
June 11th
Wet Lab
- PCR'd mCherry from NAS157
- Ran 1% Gel and purified product
Dry Lab
- Designed primers for LsR promoter
- Meeting with Dr. Sarkar
June 12th
Wet Lab
- Digested mCherry PCR product with BamHI and NotI
- Column purified mCherry and ligated into NAS152 backbone
- Transformed NAS152-mCherry into DH5alpha
- Poured 25 LB-Kan plates
Dry Lab
- Research more information about bacterial drug delivery system
- More research into biofilm project
June 14th
Wet Lab
- Fill in later....
Dry Lab
- Met with Dr. Goulian, obtained pDawn and pDusk
- Identified inaK as a surface display gene we can use
Week 3
June 18th
Wet Lab
- Miniprep pDawn and pDusk
- Test cut pDawn and pDusk with XmaI, analytical gel was correct
- Prep cut pDawn and pDusk with BamHI and NotI, gel purified
Dry Lab
- Ordered and picked up PCR purification kit from cell center
- Additional orders through cell center
- Designed primers for one of Peter's components (forgot which)
June 20
Wet Lab
- Picked 2 colonies of pDawn-mCherry, innoculated in 5 mL of LB and 50 ug/mL of Kan
- PCR purified fragments (Peter), then ran gel?
Dry Lab
- Researched DARPin binding domains and linkers
- Finalized some biobrick orders
- Finalized synthesis order (minus linker)
Week 4
June 22
Wet Lab
- Ashwin - Repeated miniprep on pDawn and did test cut
- Peter - Miniprepped pet26b and digested with BglII and EcorRI
- Avin - Miniprepped pet26b, made glycerol stock and digested with BamH1 and Not1
Dry Lab
- Avin - Finalized and sent in synthesis order (still awaiting order confirmation)
June 25
Wet Lab
- Ashwin/Avin - Column purification, ligation, and transformation of Pet26b-mCherry
- Peter -run gel of eGFP/plsr ligation
Dry Lab
- Avin - Sent in final gene synthesis order
- Mike - reviewed pDawn protocol, reviewed TetR sequences
- Peter- Order restriction enzymes from cell center
June 26
Wet Lab
- Avin - Picked colonies for Pet26b-mCherry and pJT106b
- Mike/Ashwin - Plated pJT122
Dry Lab
- Everyone - Brainstormed human practices, Wiki design
June 27
Wet Lab:
- Miniprepped pet26b-mCherry and pDawn-mCherry
- Test cut pet26b-mCherry with HindIII and ClaI (picture temporarily saved in MEAM folder of pqiao account), bands looked okay, chose C2 for remainder of experiments and made a glycerol stock, transformed both pet26b-mCherry and pDawn-mCherry into BL21(DE3)- Gold (10 ul cells, ~1ng DNA) Plated 200 ul and 20 ul
- Inoculated colonies for pJT122, pJT106b, PhyB (BBa), PIF3 (BBa), and Cph8 (BBa)
Dry Lab:
- Started exploring possible wiki designs and coding
June 28
Wet Lab:
- Miniprepped pJT122, pJT106b, PhyB, PIF3, Cph8, made glycerol stocks
- Picked colonies for pET26b-mCherry and pDawn-mCherry, innoculated, grew up, then diluted Measured OD along the way
Dry Lab:
- Set up light and dark incubators
June 29
Wet Lab:
- Tested pDawn-mCherry and pET-mCherry, pDawn-mCherry expressed mCherry only after light induction
- Nanodropped pJT122, pJT106b, PhyB, PIF3, Cph8
- Performed digest of pET-26b, eGFP, and plsr
- Spread Biobrick shipment on Amp plates (lsrR, lsrK)
Dry Lab:
- Opened Bio-Rad shipments
July 2012 Notebook
Week 5
July 2
Wet Lab:
- Transformed ho1 and pcyA BioBricks
- Peter: ligations
Dry Lab:
- Contacted labs for JT2 and pPL-PCB
- Worked on Human Practices
Week 6
Week 7
Week 8
Retrieved from "http://2012.igem.org/Team:Penn/Notebook"