2012

First thing, the morning before:

  • Pick a colony
  • Grow in 3 mL LB-Kan

Last thing, the day before: 100 µL culture into 40 mL (each w/ Kan)

  • LB
  • Lac 0.25 M
    • 20 mL LB
    • 20 mL Lac 0.5 M
  • IPTG 0.1mM
    • 20 mL LB
    • 20 mL 0.2 mM IPTG
  • Glu 0.25 M
    • 20 mL LB
    • 20 mL 0.5 M glu
  • Gal 0.25 M
    • 20 mL LB
    • 20 mL 0.5 mL gal
  • Mal 0.25 M
    • 20 mL LB
    • 20 mL 0.5 M mal
  • IPTG 0.1 mM + glu 0.25 M
    • 20 mL 0.2 mM IPTG
    • 20 mL 0.5 M glu
  • IPTG 0.1 mM + mal 0.125 M
    • 20 mL 0.2 mM IPTG
    • 10 mL LB
    • 10 mL 0.5 M mal
  • Lac 0.25 M + glu 0.25 M
    • 20 mL 0.5M lac
    • 20 mL 0.5 M glu
  • Lac 0.25 M + mal 0.125 M
    • 20 mL 0.5M lac
    • 10 mL LB
    • 10 mL 0.5 M mal
  • Gal 0.25 M + glu 0.25 M
    • 20 mL 0.5 M gal
    • 20 mL 0.5 M glu
  • Gal 0.25 M + mal 0.125 M
    • 20 mL 0.5 M gal
    • 10 mL LB
    • 10 mL 0.5 M mal

Morning of:

  • Aliquot ~4 mL each culture into plastic tubes.
  • One set per 4 tables.
  • ? save for next lab in fridge?

Media for Lac I

LB (150 mL)

LB +

  • 0.1mM IPTG (60 mL)

  • 0.5 M lac (60 mL)

  • 0.5 M glu (80 mL)

  • 0.5 M gal (60 mL)

  • 0.5 M mal (40 mL)

Make enough for Lac operon I and II:

150 mL each + 50µg/mL Kan

  • lac = 27 g

  • gal = 13.5 g

  • mal = 27 g

  • IPTG = 0.3 mL 100mM stock

200 mL glu = 18 g

Kan calculation here:

https://wahoo.nsm.umass.edu/content/media-lab-31