Team:Stockholm/15 October 2010
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==Andreas== | ==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 | ||
+ | *#BL21 pEX.SOD⋅His | ||
+ | *#BL21 pEX.nTra10⋅SOD⋅His | ||
+ | *#BL21 pEX.nTAT⋅SOD⋅His | ||
+ | *#BL21 pEX.nLMWP⋅SOD⋅His | ||
+ | *37 °C, 225 rpm | ||
+ | |||
+ | OD<sub>590</sub> measurements made every 30 min with appropriate dilution in fresh LB. | ||
+ | |||
+ | ====OD<sub>590</sub> measurements==== | ||
+ | |||
+ | {|border="1" cellpadding="1" cellspacing="0" | ||
+ | | | ||
+ | !width="50"|0 min | ||
+ | !width="50"|30 min | ||
+ | !width="50"|60 min | ||
+ | !width="50"|90 min | ||
+ | !width="50"|120 min | ||
+ | !width="50"|150 min | ||
+ | !width="50"|180 min | ||
+ | !width="50"|210 min | ||
+ | !width="50"|240 min | ||
+ | !width="50"|270 min | ||
+ | !width="50"|300 min | ||
+ | !width="50"|330 min | ||
+ | !width="50"|360 min | ||
+ | !width="50"|390 min | ||
+ | |- | ||
+ | |'''pEX.SOD⋅His''' | ||
+ | |align="center"|0.035 | ||
+ | |align="center"|0.047 | ||
+ | |align="center"|0.090 | ||
+ | |align="center"|0.180 | ||
+ | |align="center"|0.195 | ||
+ | |align="center"|0.176 | ||
+ | |align="center"|0.251 | ||
+ | |align="center"|0.177 | ||
+ | |align="center"|0.239 | ||
+ | |align="center"|0.140 | ||
+ | |align="center"|0.164 | ||
+ | |align="center"|0.205 | ||
+ | |align="center"|0.196 | ||
+ | |align="center"|0.193 | ||
+ | |- | ||
+ | |'''pEX.nTra10⋅SOD⋅His''' | ||
+ | |align="center"|0.037 | ||
+ | |align="center"|0.050 | ||
+ | |align="center"|0.092 | ||
+ | |align="center"|0.163 | ||
+ | |align="center"|0.156 | ||
+ | |align="center"|0.132 | ||
+ | |align="center"|0.175 | ||
+ | |align="center"|0.130 | ||
+ | |align="center"|0.164 | ||
+ | |align="center"|0.116 | ||
+ | |align="center"|0.155 | ||
+ | |align="center"|0.167 | ||
+ | |align="center"|0.154 | ||
+ | |align="center"|0.168 | ||
+ | |- | ||
+ | |'''pEX.nTAT⋅SOD⋅His''' | ||
+ | |align="center"|0.033 | ||
+ | |align="center"|0.046 | ||
+ | |align="center"|0.090 | ||
+ | |align="center"|0.170 | ||
+ | |align="center"|0.177 | ||
+ | |align="center"|0.161 | ||
+ | |align="center"|0.225 | ||
+ | |align="center"|0.153 | ||
+ | |align="center"|0.225 | ||
+ | |align="center"|<font style="color:red">0.045</font> | ||
+ | |align="center"|0.170 | ||
+ | |align="center"|0.162 | ||
+ | |align="center"|0.193 | ||
+ | |align="center"|0.200 | ||
+ | |- | ||
+ | |'''pEX.nLMWP⋅SOD⋅His''' | ||
+ | |align="center"|0.031 | ||
+ | |align="center"|0.042 | ||
+ | |align="center"|0.86 | ||
+ | |align="center"|0.174 | ||
+ | |align="center"|0.192 | ||
+ | |align="center"|0.178 | ||
+ | |align="center"|0.237 | ||
+ | |align="center"|0.170 | ||
+ | |align="center"|0.242 | ||
+ | |align="center"|0.137 | ||
+ | |align="center"|0.180 | ||
+ | |align="center"|0.185 | ||
+ | |align="center"|0.200 | ||
+ | |align="center"|0.200 | ||
+ | |- | ||
+ | |'''Dilution factor''' | ||
+ | |align="center"|1 | ||
+ | |align="center"|1 | ||
+ | |align="center"|1 | ||
+ | |align="center"|1 | ||
+ | |align="center"|2 | ||
+ | |align="center"|4 | ||
+ | |align="center"|5 | ||
+ | |align="center"|10 | ||
+ | |align="center"|10 | ||
+ | |align="center"|20 | ||
+ | |align="center"|20 | ||
+ | |align="center"|20 | ||
+ | |align="center"|20 | ||
+ | |align="center"|20 | ||
+ | |} | ||
+ | |||
+ | [[image:Growth_assay_plot_15oct.png|200px|thumb|right|'''Line chart of plotted OD(590) measurements from growth assay.''']] | ||
+ | |||
+ | ====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. |
Revision as of 15:53, 15 October 2010
Contents |
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
- BL21 pEX.SOD⋅His
- BL21 pEX.nTra10⋅SOD⋅His
- BL21 pEX.nTAT⋅SOD⋅His
- 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.