Difference between revisions of "Team:Nagahama/awarming"
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20 ％ (v/v) 5 × M9 salts stock solution | 20 ％ (v/v) 5 × M9 salts stock solution | ||
0.1 ％ (v/v) of CaCl2 × 2H2O stock solution (20 mg/mL) | 0.1 ％ (v/v) of CaCl2 × 2H2O stock solution (20 mg/mL) | ||
| − | + | 0.1 ％ (v/v) MgSO4 stock solution (0.12 g/mL) | |
0.03 ％ (w/v) thiamine | 0.03 ％ (w/v) thiamine | ||
0.3 ％ (w/v) agar | 0.3 ％ (w/v) agar | ||
Revision as of 12:17, 15 September 2015
Swarming Support
《We helped Tokyo_Tech.》
・Abstract We helped team Tokyo_Tech by advising these 2 about chemotaxis experiments. 1. In order to increase the chemotactic activity of E. coli, the best concentration of agar is 0.3 %. 2. It is better to directly stick the chip inside the agar and then inject the E .coli, than to place the E. coli on top of the agar Also, we gave Tokyo_Tech the raw data along with the protocol of the experiments for assaying the chemotactic activity of a wild type E.coli.
・Method 5 × M9 salts stock solution 6.4 ％ (w/v) Na2HPO4 × 7H2O 1.5 ％ (w/v) KH2PO4 0.25 ％ (w/v) NaCl 0.5 ％ (w/v) NH4Cl
M9 swarming agar 1.25 ％ (v/v) glycerol 20 ％ (v/v) 5 × M9 salts stock solution 0.1 ％ (v/v) of CaCl2 × 2H2O stock solution (20 mg/mL) 0.1 ％ (v/v) MgSO4 stock solution (0.12 g/mL) 0.03 ％ (w/v) thiamine 0.3 ％ (w/v) agar
tripton broth 1.0 ％ (w/v) tripton 1.0 ％ (w/v) NaCl
Swarming Assay We assayed chemotaxis of E. coli against cadmium and aspartic acid on soft agar, containing M9 synthetic medium, on that plate E. coli can swim. Protocol 1.E. coli JM109 was cultured at 30℃ for 12 hours with shaking (50 rpm). 2.Aliquot of the culture was spotted on the center of agar plate. 3.10 mM L-aspartic acid (40 μl) or 100 mM cadmium chloride (4 μl) was spotted on 25 mm distant from the center of the agar plate. 4.The plate was standed for 5 min at RT. 5.The plate was incubated at 30℃.
・Result and Discussion
