Team:Northwestern/07 21

Notebook

Thursday, July 21st

Tasks:

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

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