Difference between revisions of "GDSYZX-United/HUMAN PRACTICE/human practice"

 
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<ul role="menu" class="dropdown-menu">
 
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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>
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<li><a href="https://2016.igem.org/Team:GDSYZX-United/Collaborations">Collarboration</a></li>
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<li><a href="https://2016.igem.org/Team:GDSYZX-United/Attributions">Attribution</a></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>
 
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<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/project_description">Project description</a></li>
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<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>
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<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/result">Result</a></li>
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<li><a href="https://2016.igem.org/Team:GDSYZX-United/Proof">Proof</a></li>
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<li><a href="https://2016.igem.org/Team:GDSYZX-United/Demonstrate">Demonstrate</a></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>
 
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<li><a href="https://2016.igem.org/GDSYZX-United/HUMAN_PRACTICE/human_practice">Human Practice</a></li>
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<li><a href="https://2016.igem.org/Team:GDSYZX-United/HP/Silver">Silver</a></li>
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<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>
 
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<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>
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<li><a href="https://2016.igem.org/GDSYZX-United/SAFETY/safety_of_process">Safety_of_process</a></li>
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<li><a href="https://2016.igem.org/GDSYZX-United/SAFETY/safety_of_process">Safety of process</a></li>
 
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<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> ENTREPRENEURSHIP <span class="caret"></span></a>
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<a aria-expanded="false" role="button" href="https://2016.igem.org/Team:GDSYZX-United/Entrepreneurship"> ENTREPRENEURSHIP </a>
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                                    <h1><strong>Human practice</strong></h1>
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<h1><strong>Extract DNA in arabidopsis thaliana</strong></h1>
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                                    <h2><strong>Overview</strong></h2>
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<p><div class="text_font"><strong>Place:</strong>Biology Experiment Teaching Center Of Sun Yat-sen University</div></p>
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<p><div class="text_font"><strong>Date:</strong>July 25th, 2016</div></p>
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<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>
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<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>
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<p><div class="text_font"><strong>Procedure:</strong></div></p>
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<table class="table">
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<thead>
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<tr>
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<th>Experimental drugs</th>
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<th>operation</th>
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<th>Time</th>
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</tr>
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</thead>
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<tbody>
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<tr>
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<td>50-100mg Young leaves of Arabidopsis thaliana.</td>
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<td>Transfer to 2 ml centrifuge tube after grinding with grinding bar.</td>
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<td></td>
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</tr>
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<tr>
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<td>600μl 65℃ Buffer PCB  & 12μl β- mercaptoethanol</td>
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<td>Placed in 65 ℃ water after blending.</td>
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<td>25min</td>
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</tr>
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<tr>
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<td>20μl RNaseA</td>
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<td>waiting</td>
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<td>5min</td>
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</tr>
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<tr>
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<td>About 630μl chloroform</td>
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<td>Transfer  supernatant liquid to a clean 2 ml centrifuge tube after blending and following 12000 RPM centrifuge.</td>
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<td>centrifuge：5min</td>
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</tr>
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<tr>
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<td>About 400μl Buffer BD</td>
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<td>Blending for 3-5 times. </td>
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<td></td>
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</tr>
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<tr>
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<td>About 400μl anhydrous ethanol</td>
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<td>Transfer to the adsorption column after blending,and then waiting. Dump the waste liquid after centrifuging with 10000 RPM.</td>
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<td>waiting：2min<br>centrifuging：1min</td>
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</tr>
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<tr>
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<td>500μl Wash Solution</td>
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<td>Centrifuging with 10000 RPM after adding the reagent, and then dump  the waste liquid.</td>
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<td>1min</td>
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</tr>
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<tr>
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<td></td>
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<td>centrifuge with 12000 RPM.</td>
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<td>2min</td>
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</tr>
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<tr>
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<td>50μl  TE Buffer</td>
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<td>Place the adsorption column in the new 2 ml centrifuge tube, then waiting after adding reagent.</td>
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<td>wait:3min</td>
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</tr>
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<tr>
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<td></td>
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<td>centrifuge with 12000 RPM.</td>
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<td>2min</td>
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</tr>
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</tbody>
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</table>
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<p><div class="text_font"><strong>Result:</strong></div></p>
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<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>
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<p><div class="text_font"><strong>Introspection:</strong></div></p>
 
<p><div class="text_font">
 
<p><div class="text_font">
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In order to reach our goal , which is to introduce our project and amplify the impact, our team came out with some activities.</div></p>
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<ol>
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<p><div class="text_font">The efforts we made including having joined a meet-up with few universities, building up an official account, creating a questionnaire. There is a TED speech, which is provided by our members as well. If you visit our wiki, you will find a special video about our team. In this summer holiday, we completely produced a music video by every single one of our members, we filmed it, we recomposed the lines , and we gave it a name called Hey Plants,  which was based on the famous song from the Beatles.</div></p>
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<li>Understanding of the experimental requirements was not very well and had some wrong operations in the experiment.</li>
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<p><div class="text_font">Please find more details below.</div></p>
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<li>There is still many liquid residue on the grinding rod after grinding Arabidopsis thaliana leaves.</li>
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                                </div>
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<li>Not all reagents were fully blended.</li>
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                            </div>
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<li>The usage of transfer liquid gun was not normative.</li>
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</div></p>
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                                    <h2><strong>Meet-up(SUAU)</strong></h2>
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<div style="margin-bottom:30px">
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<p><div class="text_font">This summer holiday, our team has participated in a Meet-up with another iGEM team , SCAU-China from South China Agriculture University, on July the 15th. In this seminar, we aimed at several brief goals and did receive a promising result.</div></p>
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<div class="row">
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<h3><strong>TEAM FORMING</strong></h3>
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<div class="col-sm-2"></div>
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<p><div class="text_font">Having a small discussion about our project, we learned more about our process from the basic theories to the future usage of our potential products. Through  communication, we found out that there are still a few things that could be improved about our project, such as ways of experiment and methods of teamwork.</div></p>
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<div class="col-sm-8 content">
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<h3><strong>EXPERIMENTS</strong></h3>
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<p><div class="text_font">We visited their laboratories, where they were doing the experiments about their project at that time. The members of SCAU-China introduced us about some common equipment and some basic operations about molecular and synthetic biology.</div></p>
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<h3><strong>MODELING AND WIKI</strong></h3>
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<p><div class="text_font">Team members of that university recommended some common software for data modeling to us , and then they gave us an accurate instruction of the software. In addition to this, the wiki group of their team and the one of our team had a simple meeting seeking better way of designing an impressive wiki. We shared our idea with each other and got the inspiration to improve our existing plan.</div></p>
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<h3><strong>OTHERS</strong></h3>
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<p><div class="text_font">During the discussion, both of the two teams came up with some great ideas on human practice, such as activities on the Associations Opening day and building up an official account on WeChat.</div></p>
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<p><div class="text_font">Team members of SCAU-China suggested us to make a short funny video to explain our project or to introduce our team.</div></p>
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                                    <h2><strong>Official Account</strong></h2>
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<p><div class="text_font">On the most popular social app in China, called wechat, we built an official account to spread and diffuse the information about us all over the nation  about iGEM, about synthetic biology and so on.</div></p>
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<p><div class="text_font">This is our official account’s 2-dimensional bar code. More details and information about us, our project and synthetic biology are available here. Scan the code below and be a part of iGEM!</div></p>
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<div align="center">
 
<div align="center">
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<img alt="image" src="https://static.igem.org/mediawiki/2016/5/53/T--GDSYZX-United--QR_code.jpg"/>
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<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/9/94/T--GDSYZX-United--project_extract_dna1.jpg">
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<p><div class="text_font">arabidopsis thaliana</div></p>
 
</div>
 
</div>
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<p><div class="text_font">We use our official account to improve our public influence, and make people increasingly interested about us and our project. Some brilliant biological articles are released in here sometimes as well. The progress of this account  has been a great help in spreading our project and diffusing more information about synthetic biology.</div></p>
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</div>
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<p><div class="text_font">By following us on our official account, we achieve our goal and enable more people to know about our project, iGEM and synthetic biology.</div></p>
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                                    <h2><strong>Questionnaire</strong></h2>
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<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/8/80/T--GDSYZX-United--project_extract_dna2.jpg">
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<p><div class="text_font">We design a questionnaire and sent it to different quarters to complete our social research.</div></p>
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<p><div class="text_font">grinding with grinding bar</div></p>
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<p><div class="text_font">This questionnaire acquired answers to how much people know about synthetic biology and their perspective of some hot heated discussion in the field of synthetic biology. For instance, one of our questions is about transgenic technology, which is a problem that is highly controversial among Chinese people. Therefore, we ask the respondents how they understand this technology. Moreover, we intend to learn more about people's interests. By doing this questionnaire, we use the data of our result to improve our TEDxGDSYZX speech as well as other human practice projects in order to have more high-quality activities.</div></p>
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</div>
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<p><div class="text_font">And the following is our result:</div></p>
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</div>
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<p><div class="text_font">After collecting and analyzing the data, we realized that</div></p>
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<div class="col-sm-8">
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<p><div class="text_font">We hope our project can help others understand iGEM, learn more about synthetic biology and eliminate some misunderstandings on transgenic technology.</div></p>
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<div align="center">
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                                </div>
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<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/b/b1/T--GDSYZX-United--project_extract_dna3.jpg">
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                            </div>
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<p><div class="text_font">After blending with  Buffer PCB</div></p>
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                                    <h2><strong>Associations Opening Day</strong></h2>
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<div>
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<p><div class="text_font">We applied for a booth at Associations Opening Day, which is held by Guangdong Experimental high school and available to visitors all over Guangzhou, to enrich high school students' knowledge about iGEM and synthetic biology. </div></p>
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<div class="row">
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<p><div class="text_font">Through our posters and leaflets designed and manufactured before Associations Opening Day, people could be introduced to our project. A great many students are filled with curiosity and interests about our project after reading them.</div></p>
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<div class="col-sm-2"></div>
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<p><div class="text_font">A wide range of special activities are offered by us during the Associations Opening Day, including being introduced about our team, using optical microscopes, and asking questions that would be responded to as well as answered immediately. For introduction of our team, we produced posters of two kinds. One was descriptions about iGEM, synthetic biology and our project, the other was mainly constituted by some photos showing our presentation and experiment. Two of our teammates were standing close to the posters to explain the details and answer the questions.</div></p>
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<div class="col-sm-8 content">
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<p><div class="text_font">Optical microscope is one of the most common apparatus in biology experiments. We borrowed some mountings from Sun Yat-Sen University ,and took advantage of optical microscopes from Guangdong Experimental high school. </div></p>
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<div class="col-sm-6">
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<p><div class="text_font">Moreover, we prepare some simple questions about biology. If students was able to answer two of them correctly, they can get a postcard designed by our teammates.</div></p>
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<div align="center">
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<p><div class="text_font">Through the Associations Opening Day, we enable students to have some standard concepts about iGEM and a bit primary knowledge about synthetic biology.  Last but no least, we acquaint them with our project and our potential contributions.</div></p>
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<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/f/fd/T--GDSYZX-United--project_extract_dna4.jpg">
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                                </div>
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<p><div class="text_font">centrifuge</div></p>
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<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/f/f5/T--GDSYZX-United--project_extract_dna5.jpg">
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                                <div class="col-sm-8 content">
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<p><div class="text_font">result</div></p>
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                                    <h2><strong>Some legal questions</strong></h2>
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</div>
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<p><div class="text_font">According to insiders in specific fields, there are explicit safety management laws about agricultural genetically modified organisms. Forestry somehow lacks of specific regulations because of special reasons, such as the growing cycle of woods. However, forestry problems may refer to agricultural regulations. </div></p>
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</div>
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<p><div class="text_font">1.May our project refer to some legal questions?</div></p>
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</div>
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<p><div class="text_font">It can be referring to genetically modified organisms safety multi-system management. There are specific safety management laws such as regulations on Administration of Agricultural Genetically Modified Organisms Safety, Measures for the administration of Agricultural Genetically Modified Organisms Safety, Agricultural Genetically Modified Organisms Safety evaluation management approach, Implementation Regulations on the Safety of Import of Agricultural Genetically Modified Organisms, Measures for the management of Agricultural Genetically Modified Organisms processing, etc.</div></p>
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</div>
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<p><div class="text_font">2.Would the application of these anti-highlight plants steps into legal area?</div></p>
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</div>
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<p><div class="text_font">Applications, mostly generalization, particularly for merchant manufactory, merchant usage and releasing in environment, may refer to several specific law about variety certification, such as relevant regulation in Agricultural Genetically Modified Organisms Safety evaluation management approach, Measures for the examination and approval of major crop varieties which was enacted by Department of Agriculture, and Measures for the examination and approval of forest varieties which was enacted by Department of Forestry and so on. </div></p>
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</div>
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<p><div class="text_font">3.What certification should this kind of plants get before applying in actual manufacture?</div></p>
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<p><div class="text_font">Experiments, detection, and even variety judgement, are required to attain relevant certifications after passing regulation process. </div></p>
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<p><div class="text_font">Each region has its own regulation. Main forest species passing city level judgment are given certifications respectively by city Forest Variety Approval Committee, and are announced by city Forest Variety Approval Committee.</div></p>
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<p><div class="text_font">The species ,which should be certified but have not passed the judgment , should be affirmed by Forest Variety Approval Committee if needed for production. Forest Variety Approval Committee should specify using time limit and appropriate area. If generalizing outside the appropriate area which certificated by the announcement of variety certification, the plants are viewed.</div></p>
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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:

  1. Understanding of the experimental requirements was not very well and had some wrong operations in the experiment.
  2. There is still many liquid residue on the grinding rod after grinding Arabidopsis thaliana leaves.
  3. Not all reagents were fully blended.
  4. The usage of transfer liquid gun was not normative.

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arabidopsis thaliana

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grinding with grinding bar

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After blending with Buffer PCB

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centrifuge

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result