Team:Northwestern/08 08

Notebook

Monday, August 8th

Tasks:

Sara, Jordan, Shu, Tasfia

  • Ran Golden Gate reaction, transformed 5uL of each reaction
mCherry1 GFP1-mCherry2 GFP1-GFP2-mCherry3 GFP1-GFP2-GFP3-mCherry4 GFP1-GFP2-GFP3-GFP4-mCherry5
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL mC1
  • 12.5 uL H20
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL G1
  • 1 uL mC2
  • 11.5 uL H20
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL G1
  • 1 ul G2
  • 1 uL mC3
  • 10.5 uL H20
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL G1
  • 1 ul G2
  • 1 uL G3
  • 1 uL mC4
  • 9.5 uL H20
  • 2 uL BB
  • 2 uL 10X Ligase buffer
  • 1 uL ligase
  • 0.5 uL BsaI
  • 1 uL SS
  • 1 uL G1
  • 1 ul G2
  • 1 uL G3
  • 1 ul G4
  • 1 uL mC5
  • 8.5 uL H20

Paul

  • Submitted sequencing order
  • Ran gel salt test (salt+ diH2O *not autoclaved)
    • Added 1 uL of salt solution to mixture:
      • Ladder:
        • 2 uL ladder
        • 6 uL blue dye
        • 1 uL salt solution
      • Tet lin for Cas9 (~2.2 kb):
        • 4 uL DNA
        • 1 uL dye
        • 1 uL salt solution
    • Lanes 1-5 (from left to right): NEB 2-log purple ladder: 50 mM, 10 mM, 1 mM, 0.5 mM, no salt*
    • Lanes 6-10: Tet linearized for Cas9: 50 mM, 10 mM, 1 mM, 0.5 mM, no salt*
    • Should’ve added 1 uL of diH2O to control for the addition of impure water, but instead added nothing (5 uL DNA+ 1 uL dye)
  • Plated high copy Cam resistant cells from Patrick on our Cam plates, incubated

Sara

  • Made cam plates
    • 275 mL LB
    • 4.5 g bacto agar
    • Autoclave, then 275 uL Cam
  • Got 6X purple loading dye from the Jewett lab

Shu

  • Replate the transformed Gibson products in the fridge
  • Retransform the Gibson products in the freezer

Tasfia

  • Premixed sequencing rxns, “Recipe” based off of 7.26.2016 doc
  • Poured culture media for tet plates (3 mL of 5 mg/mL Tet antibiotic)

Tyler

  • Premixed sequencing rxns
  • Ordered mRFP linearizing primer
  • Blasted a few sequences for mathematical modeling, started looking into Kanamycin resistance

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