Difference between revisions of "Team:XJTLU-CHINA/Protocols Testing sensing pathway"
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<title>Testing of <i>S. aureus</i> sensing pathway</title> | <title>Testing of <i>S. aureus</i> sensing pathway</title> | ||
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<li>Dilute 1/25 in 2 x 10 ml fresh medium (30 °C). Grow until the OD<sub>600</sub> ≈ 0.4.</li> | <li>Dilute 1/25 in 2 x 10 ml fresh medium (30 °C). Grow until the OD<sub>600</sub> ≈ 0.4.</li> | ||
<li>Induce one 10 ml culture with 0.5 - 5 ng/ml nisin (typically 1 ng/ml is used) and keep the other 10 ml culture as negative control.</li> | <li>Induce one 10 ml culture with 0.5 - 5 ng/ml nisin (typically 1 ng/ml is used) and keep the other 10 ml culture as negative control.</li> | ||
| − | <li>Incubate 2 - 3 hours | + | <li>Incubate 2 - 3 hours</li> |
<ol> | <ol> | ||
<li>Aliquot both control group and experiment group into 96-well plate.</li> | <li>Aliquot both control group and experiment group into 96-well plate.</li> | ||
Latest revision as of 08:36, 31 October 2017
Testing of S. aureus sensing pathway
(Transformant with pNZ8148 harboring BBa_K2309005):
- Grow 5 ml successfully transformed bacteria culture overnight, 30 °C.
- Dilute 1/25 in 2 x 10 ml fresh medium (30 °C). Grow until the OD600 ≈ 0.4.
- Induce one 10 ml culture with 0.5 - 5 ng/ml nisin (typically 1 ng/ml is used) and keep the other 10 ml culture as negative control.
- Incubate 2 - 3 hours
- Aliquot both control group and experiment group into 96-well plate.
- Test the fluorescence under the set of 485 nm excitation and 510 nm emission.
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