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From 2010.igem.org

Contents 1 Andreas 1.1 Growth curve assay 1.1.1 Setup 1.1.2 OD590 measurements 1.1.3 Results

Andreas

Growth curve assay

Setup

Inoculated new flasks from ON cultures set 14/10:

• 250 ml E-flasks
• 25 ml LB + Amp 100 + 1 mM IPTG
• 250 μl ON culture
  1. BL21 pEX.SOD⋅His
  2. BL21 pEX.nTra10⋅SOD⋅His
  3. BL21 pEX.nTAT⋅SOD⋅His
  4. BL21 pEX.nLMWP⋅SOD⋅His
• 37 °C, 225 rpm

OD₅₉₀ measurements made every 30 min with appropriate dilution in fresh LB.

OD₅₉₀ measurements

	0 min	30 min	60 min	90 min	120 min	150 min	180 min	210 min	240 min	270 min	300 min	330 min	360 min	390 min
pEX.SOD⋅His	0.035	0.047	0.090	0.180	0.195	0.176	0.251	0.177	0.239	0.140	0.164	0.205	0.196	0.193
pEX.nTra10⋅SOD⋅His	0.037	0.050	0.092	0.163	0.156	0.132	0.175	0.130	0.164	0.116	0.155	0.167	0.154	0.168
pEX.nTAT⋅SOD⋅His	0.033	0.046	0.090	0.170	0.177	0.161	0.225	0.153	0.225	0.045	0.170	0.162	0.193	0.200
pEX.nLMWP⋅SOD⋅His	0.031	0.042	0.86	0.174	0.192	0.178	0.237	0.170	0.242	0.137	0.180	0.185	0.200	0.200
Dilution factor	1	1	1	1	2	4	5	10	10	20	20	20	20	20

Results

Results indicate no significant difference in growth between different cultures. Slightly lower curve for the nTra10-expressing clone, which is somewhat interesting, since a similar progress was seen in the canceled experiment from 14/10. At least one more experiment will be run next week, using new BL21 clones; this will show whether this is just a coincidence or not.