Difference between revisions of "Team:Goettingen/Experiments"

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<p align="center"><span style="font-size: x-large; color: #888888;"><strong>Cellulase activity screening</strong></span></p>
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<p align="center">&nbsp;</p>
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<ul>
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    <li>Streak Competent Top10 <em>E. coli</em> cells or Competent BL21 cells transformed with appropriate vector containing cellulase construct onto agar plates containing cellulose substrate.</li>
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    <li>Incubate plates at 37<sup>o</sup>C for minimum 3-4 days (timing can be extended depending on bacterial strain).</li>
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    <li>After 3-4 days look for plates having clear zone of activity around cellulose substrate.</li>
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<h2>Experiments &amp; Protocols</h2>
 
<h2>Experiments &amp; Protocols</h2>

Revision as of 09:02, 14 September 2015



LB Medium

Plasmid Extraction - using peqGOLD Plasmid Miniprep Kit I (PEQLAB Technologies)

Plasmid Extraction - using QIAprep Spin Miniprep Kit (QIAGEN)

Blunt End Ligation in pJET1.2 vector –Clone JET PCR Cloning Kit–Thermo Scientific

TOPO® Cloning protocol usingChampion™ pET Directional TOPO® Expression Kits. Thermo Fisher Scientific

PCR product purification using QIAquick® PCR Purification Kit (QIAGEN)

Plasmid transformation into chemically competent E. coli

Preparation of competent E.coli cells

Esterase activity test

PCR Gel extraction, peqGOLD Gel Extraction Kit

Electroporation of BL21 cells with pJET_RFP

Media preparation for cellulase activity screening

Cellulase activity screening

 

  • Streak Competent Top10 E. coli cells or Competent BL21 cells transformed with appropriate vector containing cellulase construct onto agar plates containing cellulose substrate.
  • Incubate plates at 37oC for minimum 3-4 days (timing can be extended depending on bacterial strain).
  • After 3-4 days look for plates having clear zone of activity around cellulose substrate.

Experiments & Protocols

Describe the experiments, research and protocols you used in your iGEM project.

What should this page contain?
  • Protocols
  • Experiments
  • Documentation of the development of your project

Inspiration