Difference between revisions of "Team:SKLMT-China/Notebook Overview"

Line 5: Line 5:
 
             <h2 class="title">Article</h2>
 
             <h2 class="title">Article</h2>
 
             <p class="content">content test</p>
 
             <p class="content">content test</p>
−
        </div>
 
−
        <div class="container">
 
−
            <div class="row">
 
−
                <ol class="breadcrumb">
 
−
                    <li><a href="#"><i class="fa fa-home">&thinsp;&thinsp;Home</i></a></li>
 
−
                    <li><a href="#"><i class="fa fa-photo">&thinsp;&thinsp;Topic</i></a></li>
 
−
                    <li class="active dropdown">
 
−
 
−
                        <!-- Catalog -->
 
−
                        <i class="dropdown-toggle" data-toggle="dropdown" aria-haspopup="true" aria-expanded="false">
 
−
                            &thinsp;&thinsp;Article
 
−
                        </i>
 
−
                        <ul class="dropdown-menu" aria-labelledby="togglea">
 
−
                            <li><a href="#">Toc 1</a></li>
 
−
                            <li><a href="#">Toc 2</a></li>
 
−
                            <li><a href="#">Toc 3</a></li>
 
−
                        </ul>
 
−
 
−
                    </li>
 
−
 
−
 
−
                </ol>
 
−
            </div>
 
 
         </div>
 
         </div>
 
     </section>
 
     </section>
−
 
+
    <section class="container">
 +
        <ol class="breadcrumb">
 +
            <li><a href="#"><i class="fa fa-home">Home</i></a></li>
 +
            <li><a href="#"><i class="fa fa-photo">Topic</i></a></li>
 +
            <li class="active"><i class="fa fa-code">Article</i></li>
 +
        </ol>
 +
    </section>
  
 
     <div class="container" id="section-2">
 
     <div class="container" id="section-2">
Line 39: Line 22:
 
                     <h4 class="panel-title">
 
                     <h4 class="panel-title">
 
                         <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">
 
                         <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">
−
                             Panel Title
+
                             Standard Protocol for Making Cells Electrocompetent
 
                         </a>
 
                         </a>
 
                     </h4>
 
                     </h4>
Line 45: Line 28:
 
                 <div id="collapseOne" class="panel-collapse collapse">
 
                 <div id="collapseOne" class="panel-collapse collapse">
 
                     <div class="panel-body">
 
                     <div class="panel-body">
−
                        Pseudomonas fluorescens Pf-5(pf5) is the only safety bacteria using in Chinese agriculture approved by Ministry of agricultural and rural affair of China . Besides ,this strain has been confirmed to be non-pathogenic to mammals. Compared with E.coli, the developed organisms , the toolkit for Pseudomonas fluorescens seems not exploit well. We are going to build a promoter library of inner promoters with various strength in pf5. We characterize the strength by a fluorescent reporter gene, firefly. An additional vector is that We hope to engineer our bacteria with the nicotine degradation gene cluster from Agrobacterium S33 into Pseudomonas fluorescens Pf-5 by rec/ET recombination , so that Pseudomonas fluorescens Pf-5 could degrade nicotine. These bacteria will be used to handle nicotine in cigarette butts or in the environment. At the same time, we would apply the promoter library to control nicotine degradation efficiency.
+
 
 
                     </div>
 
                     </div>
 
                 </div>
 
                 </div>
Line 53: Line 36:
 
                     <h4 class="panel-title">
 
                     <h4 class="panel-title">
 
                         <a data-toggle="collapse" data-parent="#accordion"  href="#collapseTwo">
 
                         <a data-toggle="collapse" data-parent="#accordion"  href="#collapseTwo">
−
                             Panel Title
+
                             Standard Protocol for Electroporation
 +
 
 
                         </a>
 
                         </a>
 
                     </h4>
 
                     </h4>
Line 59: Line 43:
 
                 <div id="collapseTwo" class="panel-collapse collapse">
 
                 <div id="collapseTwo" class="panel-collapse collapse">
 
                     <div class="panel-body">
 
                     <div class="panel-body">
−
                        Pseudomonas fluorescens Pf-5(pf5) is the only safety bacteria using in Chinese agriculture approved by Ministry of agricultural and rural affair of China . Besides ,this strain has been confirmed to be non-pathogenic to mammals. Compared with E.coli, the developed organisms , the toolkit for Pseudomonas fluorescens seems not exploit well. We are going to build a promoter library of inner promoters with various strength in pf5. We characterize the strength by a fluorescent reporter gene, firefly. An additional vector is that We hope to engineer our bacteria with the nicotine degradation gene cluster from Agrobacterium S33 into Pseudomonas fluorescens Pf-5 by rec/ET recombination , so that Pseudomonas fluorescens Pf-5 could degrade nicotine. These bacteria will be used to handle nicotine in cigarette butts or in the environment. At the same time, we would apply the promoter library to control nicotine degradation efficiency.
+
 
 
                     </div>
 
                     </div>
 
                 </div>
 
                 </div>
Line 67: Line 51:
 
                     <h4 class="panel-title">
 
                     <h4 class="panel-title">
 
                         <a data-toggle="collapse" data-parent="#accordion" href="#collapseThree">
 
                         <a data-toggle="collapse" data-parent="#accordion" href="#collapseThree">
−
                             Panel Title
+
                             Standard Protocol for Mini-Prep
 +
 
 
                         </a>
 
                         </a>
 
                     </h4>
 
                     </h4>
Line 73: Line 58:
 
                 <div id="collapseThree" class="panel-collapse collapse">
 
                 <div id="collapseThree" class="panel-collapse collapse">
 
                     <div class="panel-body">
 
                     <div class="panel-body">
−
                        abc
+
 
 
                     </div>
 
                     </div>
 
                 </div>
 
                 </div>
 
             </div>
 
             </div>
−
        </div>
+
            <div class="panel panel-default">
 +
                <div class="panel-heading">
 +
                    <h4 class="panel-title">
 +
                        <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">
  
 +
                            Standard Protocol for getting Recombineering Proficient Cells
 +
                        </a>
 +
                    </h4>
 +
                </div>
 +
                <div id="collapseOne" class="panel-collapse collapse">
 +
                    <div class="panel-body">
  
−
       
+
                    </div>
 +
                </div>
 +
            </div>
 +
            <div class="panel panel-default">
 +
                <div class="panel-heading">
 +
                    <h4 class="panel-title">
 +
                        <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">
  
−
   
+
                            Standard Protocol for Designing Oligos for Recombineering
 +
                        </a>
 +
                    </h4>
 +
                </div>
 +
                <div id="collapseOne" class="panel-collapse collapse">
 +
                    <div class="panel-body">
  
  
 +
 +
 +
 +
                    </div>
 +
                </div>
 +
            </div>
 +
            <div class="panel panel-default">
 +
                <div class="panel-heading">
 +
                    <h4 class="panel-title">
 +
                        <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">
 +
                        Standard protocol for separating a PCR-Product from its Template
 +
                        </a>
 +
                    </h4>
 +
                </div>
 +
                <div id="collapseOne" class="panel-collapse collapse">
 +
                    <div class="panel-body">
 +
 +
                    </div>
 +
                </div>
 +
            </div>
 +
           
 +
            <div class="panel panel-default">
 +
                <div class="panel-heading">
 +
                    <h4 class="panel-title">
 +
                        <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">
 +
 +
                        Standard protocol for degestion a plasmid
 +
                        </a>
 +
                    </h4>
 +
                </div>
 +
                <div id="collapseOne" class="panel-collapse collapse">
 +
                    <div class="panel-body">
 +
 +
<div class="table-wrapper">
 +
                <table class="alt">
 +
                    <thead>
 +
                        <tr>
 +
                            <th>MasterMix</th>
 +
                            <th> </th>
 +
                            <th> </th>
 +
                        </tr>
 +
                    </thead>
 +
                    <tbody>
 +
                        <tr>
 +
                            <td> Cut Smart buffer </td>
 +
                            <td>(n + 1) x </td>
 +
                            <td>0.2 µl </td>
 +
                        </tr>
 +
                        <tr>
 +
                            <td> PvuII-HF </td>
 +
                            <td>(n + 1) x </td>
 +
                            <td>0.2 µl </td>
 +
                        </tr>
 +
                        <tr>
 +
                            <td> dH2O </td>
 +
                            <td>(n + 1) x </td>
 +
                            <td>4.8 </td>
 +
                           
 +
                        </tr>
 +
                        <tr>
 +
                            <td> Total volume </td>
 +
                            <td>(n + 1) x </td>
 +
                            <td>6.0 µl </td>
 +
 +
                        </tr>
 +
                 
 +
                    </tbody>
 +
                 
 +
                </table>
 +
            </div>
 +
 +
 +
<p>• Add 6.0 µl of the Mastermix to 4.0 µl of the dissolved plasmid DNA</p>
 +
<p>• Incubate for at least 1.5 h at 37°C.</p>
 +
<p>• Load the whole mixture with loading buffer on a 0.7% agarose gel (containing EtBr) and perform electrophoresis. Take a picture.</p>
 +
 +
                    </div>
 +
                </div>
 +
            </div>
 +
 +
        </div>
 +
 +
 +
       
 +
            </article>
 +
        </div>
 +
 +
    </section>
  
 
</html>
 
</html>
  
 
{{SKLMT-China/footer}}
 
{{SKLMT-China/footer}}

Revision as of 10:05, 15 October 2018

  1. Home
  2. Topic
  3. Article

Standard Protocol for Making Cells Electrocompetent

Standard Protocol for Electroporation

Standard Protocol for Mini-Prep

Standard Protocol for getting Recombineering Proficient Cells

Standard Protocol for Designing Oligos for Recombineering

Standard protocol for separating a PCR-Product from its Template

Standard protocol for degestion a plasmid

MasterMix
Cut Smart buffer (n + 1) x 0.2 µl
PvuII-HF (n + 1) x 0.2 µl
dH2O (n + 1) x 4.8
Total volume (n + 1) x 6.0 µl

• Add 6.0 µl of the Mastermix to 4.0 µl of the dissolved plasmid DNA

• Incubate for at least 1.5 h at 37°C.

• Load the whole mixture with loading buffer on a 0.7% agarose gel (containing EtBr) and perform electrophoresis. Take a picture.

SKLMT

Useful  Links

Main Page

Special pages

Contact  us


  SKLMT.iGEM.China@gmail.com

Address


  Qingdao, Shandong


266200 Shandong University