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From 2010.igem.org

Contents 1 Andreas 1.1 Growth curve assay 1.1.1 Induction 1.1.2 OD590 measurements 1.1.3 ON cultures

Andreas

Growth curve assay

New attempt to measure growth curves for CPPs.

Induction

• 150 ml side-arm E-flasks
• 15 ml LB
• 150 μl ON culture (1:100)
  • BL21
  • ON cultures set by Mimmi 13/10
• 37 °C, 220 rpm

OD₅₉₀ measurements

	0 min	30 min	60 min	90 min	120 min	150 min	180 min	210 min	240 min	270 min
pEX.SOD⋅His	0.03	0.04	0.10	0.23	0.45	0.78	1.06	1.36	1.56	1.67
pEX.nTra10⋅SOD⋅His	0.03	0.04	0.09	0.19	0.36	0.61	0.94	1.18	1.42	1.56
pEX.nTAT⋅SOD⋅His	0.03	0.05	0.10	0.23	0.49	0.78	1.06	1.35	1.55	1.66
pEX.nLMWP⋅SOD⋅His	0.04	0.05	0.11	0.25	0.52	0.88	1.14	1.40	1.60	1.69

Experiment was canceled after 270 min as I was made aware that the spectrophotometer only makes accurate measurements up to ≈OD₅₉₀ = 0.5.

ON cultures

New ON cultures set (including two for Mimmi)

• 5 ml LB + Amp 100; 37 °C, 225 rpm
  • BL21, pEX.SOD⋅His
  • BL21, pEX.nTra10⋅SOD⋅His
  • BL21, pEX.nTAT⋅SOD⋅His
  • BL21, pEX.nLMWP⋅SOD⋅His
  • (BL21, pEX.SOD)
  • (BL21, pEX.yCCS)