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<li><a href="https://2016.igem.org/GDSYZX-United/TEAM/team_members">Team members</a></li> | <li><a href="https://2016.igem.org/GDSYZX-United/TEAM/team_members">Team members</a></li> | ||
+ | <li><a href="https://2016.igem.org/Team:GDSYZX-United/Collaborations">Collarboration</a></li> | ||
+ | <li><a href="https://2016.igem.org/Team:GDSYZX-United/Attributions">Attribution</a></li> | ||
</ul> | </ul> | ||
</li> | </li> | ||
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<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> PROJECT <span class="caret"></span></a> | <a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> PROJECT <span class="caret"></span></a> | ||
<ul role="menu" class="dropdown-menu"> | <ul role="menu" class="dropdown-menu"> | ||
− | <li><a href="https://2016.igem.org/GDSYZX-United/ | + | <li><a href="https://2016.igem.org/Team:GDSYZX-United/Description">Project description</a></li> |
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/extract _dna">Extract DNA in arabidopsis thaliana</a></li> | <li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/extract _dna">Extract DNA in arabidopsis thaliana</a></li> | ||
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/enzyme_cleavage">Enzyme cleavage</a></li> | <li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/enzyme_cleavage">Enzyme cleavage</a></li> | ||
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/preparation_arabidopsis_protoplast">Preparation of the Arabidopsis protoplast</a></li> | <li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/preparation_arabidopsis_protoplast">Preparation of the Arabidopsis protoplast</a></li> | ||
− | <li><a href="https://2016.igem.org/GDSYZX-United/ | + | <li><a href="https://2016.igem.org/Team:GDSYZX-United/Proof">Proof</a></li> |
+ | <li><a href="https://2016.igem.org/Team:GDSYZX-United/Demonstrate">Demonstrate</a></li> | ||
</ul> | </ul> | ||
</li> | </li> | ||
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<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> HUMAN PRACTICE <span class="caret"></span></a> | <a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> HUMAN PRACTICE <span class="caret"></span></a> | ||
<ul role="menu" class="dropdown-menu"> | <ul role="menu" class="dropdown-menu"> | ||
− | <li><a href="https://2016.igem.org/GDSYZX-United/ | + | <li><a href="https://2016.igem.org/Team:GDSYZX-United/HP/Silver">Silver</a></li> |
+ | <li><a href="https://2016.igem.org/Team:GDSYZX-United/HP/Gold">Gold</a></li> | ||
<li><a href="https://2016.igem.org/GDSYZX-United/HUMAN_PRACTICE/album">Album</a></li> | <li><a href="https://2016.igem.org/GDSYZX-United/HUMAN_PRACTICE/album">Album</a></li> | ||
</ul> | </ul> | ||
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<ul role="menu" class="dropdown-menu"> | <ul role="menu" class="dropdown-menu"> | ||
<li><a href="https://2016.igem.org/GDSYZX-United/SAFETY/lab_safety">Lab safety</a></li> | <li><a href="https://2016.igem.org/GDSYZX-United/SAFETY/lab_safety">Lab safety</a></li> | ||
− | <li><a href="https://2016.igem.org/GDSYZX-United/SAFETY/safety_of_process"> | + | <li><a href="https://2016.igem.org/GDSYZX-United/SAFETY/safety_of_process">Safety of process</a></li> |
</ul> | </ul> | ||
</li> | </li> | ||
<li class="dropdown"> | <li class="dropdown"> | ||
− | <a aria-expanded="false" role="button" href=" | + | <a aria-expanded="false" role="button" href="https://2016.igem.org/Team:GDSYZX-United/Entrepreneurship"> ENTREPRENEURSHIP </a> |
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<div class="ibox float-e-margins"> | <div class="ibox float-e-margins"> | ||
<div class="timeline"> | <div class="timeline"> | ||
+ | <div> | ||
+ | <div class="row"> | ||
+ | <div class="text-center"> | ||
+ | <h1><strong>Extract DNA in arabidopsis thaliana</strong></h1> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
<div class="timeline-item"> | <div class="timeline-item"> | ||
− | + | <div class="row"> | |
− | + | <div class="col-sm-2"></div> | |
− | + | <div class="col-sm-8 content"> | |
− | + | <p><div class="text_font"><strong>Place:</strong>Biology Experiment Teaching Center Of Sun Yat-sen University</div></p> | |
− | + | <p><div class="text_font"><strong>Date:</strong>July 25th, 2016</div></p> | |
− | + | <p><div class="text_font"><strong>Experimental apparatus:</strong>Grinding rods, 2 ml centrifuge tube, double panel, transfer liquid gun, adsorption column &collection tube and centrifuge.</div></p> | |
+ | <p><div class="text_font"><strong>Experimental drugs:</strong>Buffer PCB, β- mercaptoethanol, RNaseA, chloroform, Buffer BD, anhydrous ethanol, PW Solution, Wash Solution, TE Buffer.</div></p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
<div class="timeline-item"> | <div class="timeline-item"> | ||
− | + | <div class="row"> | |
− | + | <div class="col-sm-2"></div> | |
− | + | <div class="col-sm-8 content"> | |
− | + | <p><div class="text_font"><strong>Procedure:</strong></div></p> | |
+ | <table class="table"> | ||
+ | <thead> | ||
+ | <tr> | ||
+ | <th>Experimental drugs</th> | ||
+ | <th>operation</th> | ||
+ | <th>Time</th> | ||
+ | </tr> | ||
+ | </thead> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td>50-100mg Young leaves of Arabidopsis thaliana.</td> | ||
+ | <td>Transfer to 2 ml centrifuge tube after grinding with grinding bar.</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>600μl 65℃ Buffer PCB & 12μl β- mercaptoethanol</td> | ||
+ | <td>Placed in 65 ℃ water after blending.</td> | ||
+ | <td>25min</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>20μl RNaseA</td> | ||
+ | <td>waiting</td> | ||
+ | <td>5min</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>About 630μl chloroform</td> | ||
+ | <td>Transfer supernatant liquid to a clean 2 ml centrifuge tube after blending and following 12000 RPM centrifuge.</td> | ||
+ | <td>centrifuge:5min</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>About 400μl Buffer BD</td> | ||
+ | <td>Blending for 3-5 times. </td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>About 400μl anhydrous ethanol</td> | ||
+ | <td>Transfer to the adsorption column after blending,and then waiting. Dump the waste liquid after centrifuging with 10000 RPM.</td> | ||
+ | <td>waiting:2min<br>centrifuging:1min</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>500μl Wash Solution</td> | ||
+ | <td>Centrifuging with 10000 RPM after adding the reagent, and then dump the waste liquid.</td> | ||
+ | <td>1min</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>centrifuge with 12000 RPM.</td> | ||
+ | <td>2min</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>50μl TE Buffer</td> | ||
+ | <td>Place the adsorption column in the new 2 ml centrifuge tube, then waiting after adding reagent.</td> | ||
+ | <td>wait:3min</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>centrifuge with 12000 RPM.</td> | ||
+ | <td>2min</td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="timeline-item" style="margin-bottom:30px"> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-2"></div> | ||
+ | <div class="col-sm-8 content"> | ||
+ | <p><div class="text_font"><strong>Result:</strong></div></p> | ||
+ | <p><div class="text_font">The target DNA can be extracted successfully in most of the groups, but they have very low concentration and many impurities.</div></p> | ||
+ | <p><div class="text_font"><strong>Introspection:</strong></div></p> | ||
<p><div class="text_font"> | <p><div class="text_font"> | ||
− | + | <ol> | |
− | + | <li>Understanding of the experimental requirements was not very well and had some wrong operations in the experiment.</li> | |
− | + | <li>There is still many liquid residue on the grinding rod after grinding Arabidopsis thaliana leaves.</li> | |
− | + | <li>Not all reagents were fully blended.</li> | |
− | + | <li>The usage of transfer liquid gun was not normative.</li> | |
− | + | </ol> | |
− | + | </div></p> | |
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | <div style="margin-bottom:30px"> | |
− | + | <div class="row"> | |
− | + | <div class="col-sm-2"></div> | |
− | + | <div class="col-sm-8 content"> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
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− | <div | + | |
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<div align="center"> | <div align="center"> | ||
− | <img alt="image" src="https://static.igem.org/mediawiki/2016/ | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/9/94/T--GDSYZX-United--project_extract_dna1.jpg"> |
+ | <p><div class="text_font">arabidopsis thaliana</div></p> | ||
</div> | </div> | ||
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | <div style="margin-bottom:30px"> | |
− | + | <div class="row"> | |
− | <div | + | <div class="col-sm-2"></div> |
− | + | <div class="col-sm-8 content"> | |
− | + | <div class="col-sm-4"> | |
− | + | <div align="center"> | |
− | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/8/80/T--GDSYZX-United--project_extract_dna2.jpg"> | |
− | + | <p><div class="text_font">grinding with grinding bar</div></p> | |
− | + | </div> | |
− | + | </div> | |
− | + | <div class="col-sm-8"> | |
− | + | <div align="center"> | |
− | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/b/b1/T--GDSYZX-United--project_extract_dna3.jpg"> | |
− | + | <p><div class="text_font">After blending with Buffer PCB</div></p> | |
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | <div> | |
− | + | <div class="row"> | |
− | + | <div class="col-sm-2"></div> | |
− | + | <div class="col-sm-8 content"> | |
− | + | <div class="col-sm-6"> | |
− | + | <div align="center"> | |
− | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/f/fd/T--GDSYZX-United--project_extract_dna4.jpg"> | |
− | + | <p><div class="text_font">centrifuge</div></p> | |
− | + | </div> | |
− | + | </div> | |
− | <div | + | <div class="col-sm-6"> |
− | + | <div align="center"> | |
− | + | <img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/f/f5/T--GDSYZX-United--project_extract_dna5.jpg"> | |
− | + | <p><div class="text_font">result</div></p> | |
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | </div> | |
− | + | ||
− | + | ||
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</div> | </div> | ||
</div> | </div> |
Latest revision as of 03:01, 12 October 2016
Extract DNA in arabidopsis thaliana
Place:Biology Experiment Teaching Center Of Sun Yat-sen University
Date:July 25th, 2016
Experimental apparatus:Grinding rods, 2 ml centrifuge tube, double panel, transfer liquid gun, adsorption column &collection tube and centrifuge.
Experimental drugs:Buffer PCB, β- mercaptoethanol, RNaseA, chloroform, Buffer BD, anhydrous ethanol, PW Solution, Wash Solution, TE Buffer.
Procedure:
Experimental drugs | operation | Time |
---|---|---|
50-100mg Young leaves of Arabidopsis thaliana. | Transfer to 2 ml centrifuge tube after grinding with grinding bar. | |
600μl 65℃ Buffer PCB & 12μl β- mercaptoethanol | Placed in 65 ℃ water after blending. | 25min |
20μl RNaseA | waiting | 5min |
About 630μl chloroform | Transfer supernatant liquid to a clean 2 ml centrifuge tube after blending and following 12000 RPM centrifuge. | centrifuge:5min |
About 400μl Buffer BD | Blending for 3-5 times. | |
About 400μl anhydrous ethanol | Transfer to the adsorption column after blending,and then waiting. Dump the waste liquid after centrifuging with 10000 RPM. | waiting:2min centrifuging:1min |
500μl Wash Solution | Centrifuging with 10000 RPM after adding the reagent, and then dump the waste liquid. | 1min |
centrifuge with 12000 RPM. | 2min | |
50μl TE Buffer | Place the adsorption column in the new 2 ml centrifuge tube, then waiting after adding reagent. | wait:3min |
centrifuge with 12000 RPM. | 2min |
Result:
The target DNA can be extracted successfully in most of the groups, but they have very low concentration and many impurities.
Introspection:
- Understanding of the experimental requirements was not very well and had some wrong operations in the experiment.
- There is still many liquid residue on the grinding rod after grinding Arabidopsis thaliana leaves.
- Not all reagents were fully blended.
- The usage of transfer liquid gun was not normative.
arabidopsis thaliana
grinding with grinding bar
After blending with Buffer PCB
centrifuge
result