Difference between revisions of "Team:ShanghaitechChina/Notebook"

 
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<font color="red">The following steps work for the experiments which conduct the characterization of biofilm via solution.</font>
 
<font color="red">The following steps work for the experiments which conduct the characterization of biofilm via solution.</font>
 
<h4><b>Part I  Preparation for the Characterization: The activation of the bacteria.</b></h4>
 
<h4><b>Part I  Preparation for the Characterization: The activation of the bacteria.</b></h4>
Take the CsgA - Histag mutant E.coli as a specific example.<p></p>
+
Take the CsgA - Histag mutant <i>E.coli</i> as a specific example.<p></p>
 
i.First, prepare a aseptic condition with the usage of Bunsen burner, which is able to  achieve sterilization in a limited extent.<p></p>
 
i.First, prepare a aseptic condition with the usage of Bunsen burner, which is able to  achieve sterilization in a limited extent.<p></p>
 
ii.Take 5ml Luria-Bertani liquid medium into the centrifuge tube with both the LB medium and the centrifuge tube sterilized.<p></p>
 
ii.Take 5ml Luria-Bertani liquid medium into the centrifuge tube with both the LB medium and the centrifuge tube sterilized.<p></p>
 
iii.Take 34mg/ml Chloramphenicol which is kept in minus 20 centigrade and diluted 10 times in the 5ml LB liquid medium.<p></p>
 
iii.Take 34mg/ml Chloramphenicol which is kept in minus 20 centigrade and diluted 10 times in the 5ml LB liquid medium.<p></p>
iv. Take the CsgA - Histag mutant E.coli which is kept in minus 80 centigrade into the centrifuge tube with the Chloramphenicol added. <p></p>
+
iv. Take the CsgA - Histag mutant <i>E.coli</i> which is kept in minus 80 centigrade into the centrifuge tube with the Chloramphenicol added. <p></p>
 
<h4><b>Part II Characterization of Biofilm</b></h4>
 
<h4><b>Part II Characterization of Biofilm</b></h4>
i.Take out the CsgA - Histag mutant E.coli which is activated in Part I.<p></p>
+
i.Take out the CsgA - Histag mutant <i>E.coli</i> which is activated in Part I.<p></p>
ii. Centrifuge the mutant E.coli with 5000 g for 1minute and leave the supermatant.<p></p>
+
ii. Centrifuge the mutant <i>E.coli</i> with 5000 g for 1minute and leave the supermatant.<p></p>
 
iii. Prepare the liquid medium as the following components.<p></p>
 
iii. Prepare the liquid medium as the following components.<p></p>
 
<p><b>Treat and Control group:</b></p>
 
<p><b>Treat and Control group:</b></p>
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<font color="red">The following steps work for the experiments which conduct the characterization of biofilm via CongoRed Plates.</font>
 
<font color="red">The following steps work for the experiments which conduct the characterization of biofilm via CongoRed Plates.</font>
 
<h4><b>Part I  Preparation for the Characterization: The activation of the bacteria.</b></h4>
 
<h4><b>Part I  Preparation for the Characterization: The activation of the bacteria.</b></h4>
Take the CsgA - Histag mutant E.coli as a specific example.<p></p>
+
Take the CsgA - Histag mutant <i>E.coli</i> as a specific example.<p></p>
 
i.First, prepare a aseptic condition with the usage of Bunsen burner, which is able to  achieve sterilization in a limited extent.<p></p>
 
i.First, prepare a aseptic condition with the usage of Bunsen burner, which is able to  achieve sterilization in a limited extent.<p></p>
 
ii.Take 5ml Luria-Bertani liquid medium into the centrifuge tube with both the LB medium and the centrifuge tube sterilized.<p></p>
 
ii.Take 5ml Luria-Bertani liquid medium into the centrifuge tube with both the LB medium and the centrifuge tube sterilized.<p></p>
 
iii.Take 34mg/ml Chloramphenicol which is kept in minus 20 centigrade and diluted 10 times in the 5ml LB liquid medium.<p></p>
 
iii.Take 34mg/ml Chloramphenicol which is kept in minus 20 centigrade and diluted 10 times in the 5ml LB liquid medium.<p></p>
iv. Take the CsgA - Histag mutant E.coli which is kept in minus 80 centigrade into the centrifuge tube with the Chloramphenicol added. <p></p>
+
iv. Take the CsgA - Histag mutant <i>E.coli</i> which is kept in minus 80 centigrade into the centrifuge tube with the Chloramphenicol added. <p></p>
 
<h4><b>Part II Preparation for the CongoRed Plates.</b></h4>
 
<h4><b>Part II Preparation for the CongoRed Plates.</b></h4>
 
Make  the CongoRed Plates with the components as the following:<p></p>
 
Make  the CongoRed Plates with the components as the following:<p></p>
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vii. Brilliant Blue G250 0.5mg/100ml;<p></p>
 
vii. Brilliant Blue G250 0.5mg/100ml;<p></p>
 
<h4><b>Part III  Characterization of biofilm in CongoRed Plates.</b></h4>
 
<h4><b>Part III  Characterization of biofilm in CongoRed Plates.</b></h4>
Take the activated CsgA-Histag mutant E.coli in small amount into the plates and cultivate them for 48 hours and then the CongoRed plates will show red biofilm vividly if the experiment succeed.<p></p>
+
Take the activated CsgA-Histag mutant <i>E.coli</i> in small amount into the plates and cultivate them for 48 hours and then the CongoRed plates will show red biofilm vividly if the experiment succeed.<p></p>
 
<font color="red">The following steps work for the experiments which can dye the biofilm with the crystal violet.</font><p></p>
 
<font color="red">The following steps work for the experiments which can dye the biofilm with the crystal violet.</font><p></p>
 
         i.Add 400 ul 0.1% crystal violet dye to the medium, and hatch for 10 to15 minutes.<p></p>
 
         i.Add 400 ul 0.1% crystal violet dye to the medium, and hatch for 10 to15 minutes.<p></p>

Latest revision as of 21:56, 19 October 2016

igem2016:ShanghaiTech