Jordan

• Transformed J04550 RFP part to amplify for gRNA construct
  • Resuspended Kit Plate 4, well 2H in 10 uL of water, transformed 2 uL of that into 50 uL comp cells
  • Plated 50 and 100 uL
• Transformed competent cell test
  • 7 test conditions: 50, 20, 10, 5, and .5 pg/uL of RFP plasmid (part J04550) and 1.11 ng/ul and 8.9 ng/ul of pSB1C3 + tet plasmid
  • Transformed 1 ul of each
  • Did two replicate plates of each

Michelle

• PCR INP part out of the iGEM registry
• 10 uL dH₂O, pipet up and down, sit 5 min
• Transformed INP biobrick
• Transformed using 1 uL of diluted DNA from plate
• Followed bootcamp protocol except:
  • 50 mL competent cells instead of 100 mL
  • 450 uL SOC instead of 900 uL
  • Used 108.3 ng/uL DNA straight from the iGEM plate
• Plated 100 uL
• Ran gel on PCR product

Paul

• Planned gRNA assembly procedure
• Selected storage plasmids for use in Golden Gate
• Transformed mRFP device:
  • Transformed using 1 uL of diluted DNA from plate
  • Followed bootcamp protocol except:
    • 50 mL competent cells instead of 100 mL
    • 450 uL SOC instead of 900 uL
    • Used 108.3 ng/uL DNA straight from the iGEM plate
  • Plated 100 uL

Sam

• Went to the products showcase thing to ask for free stuff
• Found more antibiotic experts

Sara

• Nanodropped the Gibson Tet Cas9 product - 1730 ng/uL
• Diluted the Gibson product to 50 ng/uL
  • 1.16 uL Gibson product
  • 38.84 uL NF water
  • 40 uL in total
• Restriction digestion of Gibson
  • 5 uL 10X NE buffer 4
  • 1 uL EcoRi HF
  • 20 uL 50 ng/uL Diluted Gibson (1 ug)
  • 25 uL water
  • 50 uL in total
• Ran a gel of the restriction digest with Shu
  • 8 uL ladder
  • 6 uL DNA
    • 2.5 uL restricted DNA (50 ng)
    • 2.5 uL NF water
    • 1 uL loading dye

Shu

• Ran a gel of the restriction digest with Sara
• Sorting out the things we received from IDT today
• Read Golden Gate papers
• Looked into the traveling grant

Tasfia

• PCR’ed INP and ran a gel for product
  • 2.5 uL 10X PCR buffer
  • 0.5 uL 10 mM dNTPs
  • 0.5 uL INP-fwd at 10 uM
  • 0.5 uL INP-rev at 10 uM
  • 0.5 uL template (BB part, 2.4 kb, so 2 min 24s of elongation)
  • 1 hr at 50oC Total of 20 uL
• Transformed INP
• Transformed using 1 uL of diluted DNA from plate
• Followed bootcamp protocol except:
  • 50 mL competent cells instead of 100 mL
  • 450 uL SOC instead of 900 uL
  • Used 108.3 ng/uL DNA straight from the iGEM plate
• Plated 100 uL
• Worked on presentation slideshow