Team:Ehime-Japan/Modeling

From 2012.igem.org

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<p>Method<p/>
<p>Method<p/>
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E.coli (pIJ106b, pJT122, plPCB) is cultivated in TY culture medium for 7 hours. It was precipitated by centrifugation and washed with 1 % NaClaq.
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400 &mu;L of 1% NaCl was added to the E.coli sample and protein synthesis was stopped by addition of tetracyclin at the concentration of 100 &mu;g/mL.
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An aliquot from each sample was plated on agar and cultured overnight.
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The solution was exposed to blue LED light, and we took a color picture every 1 hour for 6 hours.
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By using the ImageJ software, we measured the intensity of light from GFP.
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Revision as of 03:10, 27 September 2012

Ehime-Japan iGEM Team: Welcome


Introduction

In our experiment of the E.co-mail, the important thing is to degrade GFP-Lon-tag more rapidly. GFP is usually stable and has a long half–life. So, we thought of how to measure the reaction rate.

Method

E.coli (pIJ106b, pJT122, plPCB) is cultivated in TY culture medium for 7 hours. It was precipitated by centrifugation and washed with 1 % NaClaq. 400 μL of 1% NaCl was added to the E.coli sample and protein synthesis was stopped by addition of tetracyclin at the concentration of 100 μg/mL. An aliquot from each sample was plated on agar and cultured overnight. The solution was exposed to blue LED light, and we took a color picture every 1 hour for 6 hours. By using the ImageJ software, we measured the intensity of light from GFP.

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