Team:Penn/Notebook

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

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

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)

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)

Update for June 25 Wet Lab Ashwin/Avin - Column purification, ligation, and transformation of Pet26b-mCherry Mike - Peter -run gel of eGFP/plsr ligation Nikita - Dry Lab Avin - Sent in final gene synthesis order Mike - reviewed pDawn protocol, reviewed TetR sequences Peter- Order restriction enzymes from cell center