20.109(F18): Prep notes for M2

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20.109(F18): Laboratory Fundamentals of Biological Engineering

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Fall 2018 schedule        FYI        Assignments        Homework        Class data        Communication
       1. Measuring genomic instability        2. Modulating metabolism        3. Engineering biomaterials              


M2D1

per team:

  • 100 μL nuclease-free H2O2
  • ~ 2 μg pdCas9
  • not aliquoted: enzyme stocks, buffer stocks

M2D2

per team:

  • 25 μL 6X loading dye
  • not aliquoted: 1 kb ladder stock, electrophoresis buffer

M2D3

per team:

  • 750 μL nuclease-free H2O2
  • not aliquoted: nuclease-free H2O2, pgRNA vector, gRNA reverse primer, Q5 Master Mix

M2D4

per team:

  • none

M2D5

per team:
From Qiagen mini-prep kit

  • 550 μL buffer P1
  • 550 μL buffer P2
  • 750 μL buffer N3
  • 1 mL buffer PB
  • 1.6 mL buffer PE
  • 70 μL nuclease-free H2O2, pH = 8
  • 1 mL SOC

not aliquoted: gRNA sequencing primers

M2D6

per team:

  • none

M2D7

per team:

  • 45 mL LB broth
  • 50 μL chloramphenicol stock
  • 50 μL ampicillin stock

M2D8

per team:

  • each team uses either Ethanol kit OR Acetate kit reagents

From Ethanol kit (store at 4°C, except ethanol probe at -20°C)

  • 20 mL LB broth
  • 5 mL ethanol assay buffer
  • 83 μL ethanol probe
  • 83 μL ethanol enzyme

From Acetate kit (store at -20°C, except LB at 4°C)

  • 10 mL LB broth
  • 5.6 mL acetate assay buffer
  • 86.4 μL acetate probe
  • 86.4 μL acetate substrate
  • 86.4 μL ATP
  • 86.4 μL acetate enzyme