Difference between revisions of "GDSYZX-United/NOTEBOOK/day note"

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<nav class="navbar navbar-fixed-top navbar-inverse" role="navigation">
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<!-- <div class="navbar-header">
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<a href="#" class="navbar-brand">GDSYZX-United</a>
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<a aria-expanded="false" role="button" href="https://2016.igem.org/Team:GDSYZX-United"> HOME </a>
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<li><a href="https://2016.igem.org/GDSYZX-United/TEAM/team_members">Team members</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>
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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/GDSYZX-United/PROJECT/extract _dna">Extract DNA in arabidopsis thaliana</a></li>
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<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/enzyme_cleavage">Enzyme cleavage</a></li>
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<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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<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> NOTEBOOK <span class="caret"></span></a>
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    <li><a href="https://2016.igem.org/GDSYZX-United/NOTEBOOK/day_note">Day notes</a></li>
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    <li><a href="https://2016.igem.org/GDSYZX-United/NOTEBOOK/procedure_record">Procedure record</a></li>
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    <li><a href="https://2016.igem.org/GDSYZX-United/NOTEBOOK/protocol">Protocol</a></li>
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</ul>
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<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/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>
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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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<li><a href="#">1</a></li>
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<li><a href="#">2</a></li>
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<li><a href="#">3</a></li>
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<li><a href="#">4</a></li>
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        <div class="row animated fadeInRight">
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                                    <h1>Day note</h1>
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                                </div>
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                            </div>
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                        </div>
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                        <div class="timeline-item">
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                            <div class="row">
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                                <div class="col-xs-3 date">
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                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.7.31</div></p>
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<small class="text-navy"><p><div class="text_font">9:00</div></small></p>
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                                </div>
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                                <div class="col-xs-8 content">
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                                    <div class="m-b-xs text_font"><strong>Extract genome from Arabidopsis Thaliana</strong></div>
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                                    <p><div class="text_font">The genome has been degraded greatly,possibly because we didn’t grind the leaves in very low temperature to inhibit the nuclease.We have to do it again.</div></p>
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                                </div>
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                            </div>
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                        </div>
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                        <div class="timeline-item">
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                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.7.31</div></p>
 +
<small class="text-navy"><p><div class="text_font">13:30</div></small></p>
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                                </div>
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                                <div class="col-xs-8 content">
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                                    <div class="m-b-xs text_font"><strong>Extract genome from Arabidopsis Thaliana</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We extracted it the genome successfully.But the genome has still been degraded a little. (Since we don’t have liquid nitrogen in our lab,so we grinded  the  leaves in an ice box. )We will use the genome to do the following PCR.
 +
                                    </div></p>
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                                </div>
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                            </div>
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                        </div>
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                        <div class="timeline-item">
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                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.1</div></p>
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<small class="text-navy"><p><div class="text_font">9:00</div></small></p>
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                                </div>
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                                <div class="col-xs-8 content">
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                                    <div class="m-b-xs text_font"><strong>We use PCR to generate and to amplify our target DNA fragments:promotercop1,promoter phyB,promoterpif1,gene hhl1,gene pHhl,gene flu.</strong>
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                                    </div>
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                                    <p><div class="text_font">
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                                        We failed to get all our target DNA fragments .We will check the primers and the PCR program ,and then do it again.
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                                    </div></p>
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                                </div>
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                            </div>
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                        </div>
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                        <div class="timeline-item">
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                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.2</div></p>
 +
<small class="text-navy"><p><div class="text_font">9:00</div></small></p>
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                                </div>
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                                <div class="col-xs-8 content">
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                                    <div class="m-b-xs text_font"><strong>Do the PCR again to get our target DNA fragments.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We PCR gene pHhl1,gene hhl1,gene flu successfully.
 +
                                        We will PCR cop1 together with pif1,phyB tomorrow.
 +
                                    </div></p>
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                                </div>
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                            </div>
 +
                        </div>
 +
                        <div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.3</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">9:00</div></small></p>
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                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We use PCR to get cop1,phyB and pif1</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        In the first PCR we get cop 1 and pif1,we get higher concentration of phyB in the second time PCR.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.3</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">13:30</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We use a kit box to purify our PCR products,so we can get our genes.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.3</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the afternoon</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We cut our DNA fragments using restriction enzymes:BsaⅠ,PstⅠ,
 +
EcorⅠ.After this process ,the gene fragments will have one overhang at both ends.The overhangs will help them splice other fragments. </strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">9:00</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We purify our genes from the restriction enzyme systems.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the morning</div></small></p>
 +
                                </div>
 +
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We cut our plasmids psb3c1 with BsaⅠ.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the morning</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We splice our DNA fragments to get the parts we want using
 +
a kit box.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">13:30</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We purify our parts using a kit box.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the afternoon</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We cut our parts with BsaⅠto
 +
make sure there will be no loops in the part.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the afternoon</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We purify the parts.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the afternoon</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We splice our parts with plasmids.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the afternoon</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We prepare LB broth.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">19:00</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We transform our plasmids into DH5α.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.4</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the evening</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We inoculate Ecoli DH5α on LB plates and in media.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.5</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">13:00</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We check the growth of Ecoli on LB plates.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        No conlony was observed.
 +
So we will use the Ecoli in media to do the following experiments.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.5</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the afternoon</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We extract plasmids from the solution.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.6</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">9:00</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We prepare Arabidopsis protoplasts.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        The concentration of protoplasts  is too low,this is because we chose the wrong filter paper.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.6</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">13:00</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We prepare protoplasts again.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.The concentration is much higher,and there is much more sediment of protoplast after centrifuging.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.6</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the afternoon</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We transfect our plasmids into the protoplasts，and incubate
 +
the protoplasts in the following light intensity (lux):0,3960,7920,11880.
 +
</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.7</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">9:00</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We extract RNA from protoplasts using a kit box.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We did it.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.7</div></p>
 +
                                    <small class="text-navy"><p><div class="text_font">in the morning</div></small></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>We use semiquantitive RT-PCR
 +
to amplify hhl1 to see how much hhl1 is transcripted into mRNA. 
 +
</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We failed to get any hhl1.  There may be something wrong with the process of transformation.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
<div class="timeline-item">
 +
                            <div class="row">
 +
                                <div class="col-xs-3 date">
 +
                                    <i class="fa fa-file-text"></i><p><div class="text_font"> 2016.8.10</div></p>
 +
                                </div>
 +
                                <div class="col-xs-8 content">
 +
                                    <div class="m-b-xs text_font"><strong>PCR our target DNA fragments:cop1,pif1,phyB,hhl1,pHhl1-hhl1,flu.</strong>
 +
                                    </div>
 +
                                    <p><div class="text_font">
 +
                                        We will check the result tomorrow.
 +
                                    </div></p>
 +
                                </div>
 +
                            </div>
 +
                        </div>
 +
                    </div>
 +
                </div>
 +
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Revision as of 14:14, 30 September 2016

Day note

2016.7.31

9:00

Extract genome from Arabidopsis Thaliana

The genome has been degraded greatly,possibly because we didn’t grind the leaves in very low temperature to inhibit the nuclease.We have to do it again.

2016.7.31

13:30

Extract genome from Arabidopsis Thaliana

We extracted it the genome successfully.But the genome has still been degraded a little. (Since we don’t have liquid nitrogen in our lab,so we grinded the leaves in an ice box. )We will use the genome to do the following PCR.

2016.8.1

9:00

We use PCR to generate and to amplify our target DNA fragments:promotercop1,promoter phyB,promoterpif1,gene hhl1,gene pHhl,gene flu.

We failed to get all our target DNA fragments .We will check the primers and the PCR program ,and then do it again.

2016.8.2

9:00

Do the PCR again to get our target DNA fragments.

We PCR gene pHhl1,gene hhl1,gene flu successfully. We will PCR cop1 together with pif1,phyB tomorrow.

2016.8.3

9:00

We use PCR to get cop1,phyB and pif1

In the first PCR we get cop 1 and pif1,we get higher concentration of phyB in the second time PCR.

2016.8.3

13:30

We use a kit box to purify our PCR products,so we can get our genes.

We did it.

2016.8.3

in the afternoon

We cut our DNA fragments using restriction enzymes:BsaⅠ,PstⅠ, EcorⅠ.After this process ,the gene fragments will have one overhang at both ends.The overhangs will help them splice other fragments.

We did it.

2016.8.4

9:00

We purify our genes from the restriction enzyme systems.

We did it.

2016.8.4

in the morning

We cut our plasmids psb3c1 with BsaⅠ.

We did it.

2016.8.4

in the morning

We splice our DNA fragments to get the parts we want using a kit box.

We did it.

2016.8.4

13:30

We purify our parts using a kit box.

We did it.

2016.8.4

in the afternoon

We cut our parts with BsaⅠto make sure there will be no loops in the part.

We did it.

2016.8.4

in the afternoon

We purify the parts.

We did it.

2016.8.4

in the afternoon

We splice our parts with plasmids.

We did it.

2016.8.4

in the afternoon

We prepare LB broth.

We did it.

2016.8.4

19:00

We transform our plasmids into DH5α.

We did it.

2016.8.4

in the evening

We inoculate Ecoli DH5α on LB plates and in media.

We did it.

2016.8.5

13:00

We check the growth of Ecoli on LB plates.

No conlony was observed. So we will use the Ecoli in media to do the following experiments.

2016.8.5

in the afternoon

We extract plasmids from the solution.

We did it.

2016.8.6

9:00

We prepare Arabidopsis protoplasts.

The concentration of protoplasts is too low,this is because we chose the wrong filter paper.

2016.8.6

13:00

We prepare protoplasts again.

We did it.The concentration is much higher,and there is much more sediment of protoplast after centrifuging.

2016.8.6

in the afternoon

We transfect our plasmids into the protoplasts，and incubate the protoplasts in the following light intensity (lux):0,3960,7920,11880.

We did it.

2016.8.7

9:00

We extract RNA from protoplasts using a kit box.

We did it.

2016.8.7

in the morning

We use semiquantitive RT-PCR to amplify hhl1 to see how much hhl1 is transcripted into mRNA.

We failed to get any hhl1. There may be something wrong with the process of transformation.

2016.8.10

PCR our target DNA fragments:cop1,pif1,phyB,hhl1,pHhl1-hhl1,flu.

We will check the result tomorrow.

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Day note

2016.7.31

9:00
</small>

                               </div>
Extract genome from Arabidopsis Thaliana

The genome has been degraded greatly,possibly because we didn’t grind the leaves in very low temperature to inhibit the nuclease.We have to do it again.

                           </div>
                       </div>

2016.7.31

13:30
</small>

                               </div>
Extract genome from Arabidopsis Thaliana 

                                       We extracted it the genome successfully.But the genome has still been degraded a little. (Since we don’t have liquid nitrogen in our lab,so we grinded  the  leaves in an ice box. )We will use the genome to do the following PCR.


                           </div>
                       </div>

2016.8.1

9:00
</small>

                               </div>
We use PCR to generate and to amplify our target DNA fragments:promotercop1,promoter phyB,promoterpif1,gene hhl1,gene pHhl,gene flu. 

                                       We failed to get all our target DNA fragments .We will check the primers and the PCR program ,and then do it again.


                           </div>
                       </div>

2016.8.2

9:00
</small>

                               </div>
Do the PCR again to get our target DNA fragments. 

                                       We PCR gene pHhl1,gene hhl1,gene flu successfully.
                                       We will PCR cop1 together with pif1,phyB tomorrow.


                           </div>
                       </div>

2016.8.3

9:00
</small>

                               </div>
We use PCR to get cop1,phyB and pif1 

                                       In the first PCR we get cop 1 and pif1,we get higher concentration of phyB in the second time PCR.


                           </div>
                       </div>

2016.8.3

13:30
</small>

                               </div>
We use a kit box to purify our PCR products,so we can get our genes. 

                                       We did it.


                           </div>
                       </div>

2016.8.3

in the afternoon
</small>

                               </div>
We cut our DNA fragments using restriction enzymes:BsaⅠ,PstⅠ,

EcorⅠ.After this process ,the gene fragments will have one overhang at both ends.The overhangs will help them splice other fragments. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

9:00
</small>

                               </div>
We purify our genes from the restriction enzyme systems. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

in the morning
</small>

                               </div>
We cut our plasmids psb3c1 with BsaⅠ. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

in the morning
</small>

                               </div>
We splice our DNA fragments to get the parts we want using

a kit box. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

13:30
</small>

                               </div>
We purify our parts using a kit box. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

in the afternoon
</small>

                               </div>
We cut our parts with BsaⅠto

make sure there will be no loops in the part. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

in the afternoon
</small>

                               </div>
We purify the parts. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

in the afternoon
</small>

                               </div>
We splice our parts with plasmids. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

in the afternoon
</small>

                               </div>
We prepare LB broth. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

19:00
</small>

                               </div>
We transform our plasmids into DH5α. 

                                       We did it.


                           </div>
                       </div>

2016.8.4

in the evening
</small>

                               </div>
We inoculate Ecoli DH5α on LB plates and in media. 

                                       We did it.


                           </div>
                       </div>

2016.8.5

13:00
</small>

                               </div>
We check the growth of Ecoli on LB plates. 

                                       No conlony was observed.

So we will use the Ecoli in media to do the following experiments. 

                           </div>
                       </div>

2016.8.5

in the afternoon
</small>

                               </div>
We extract plasmids from the solution. 

                                       We did it.


                           </div>
                       </div>

2016.8.6

9:00
</small>

                               </div>
We prepare Arabidopsis protoplasts. 

                                       The concentration of protoplasts  is too low,this is because we chose the wrong filter paper. 


                           </div>
                       </div>

2016.8.6

13:00
</small>

                               </div>
We prepare protoplasts again. 

                                       We did it.The concentration is much higher,and there is much more sediment of protoplast after centrifuging.


                           </div>
                       </div>

2016.8.6

in the afternoon
</small>

                               </div>
We transfect our plasmids into the protoplasts，and incubate

the protoplasts in the following light intensity (lux):0,3960,7920,11880. 

                                       We did it.


                           </div>
                       </div>

2016.8.7

9:00
</small>

                               </div>
We extract RNA from protoplasts using a kit box. 

                                       We did it.


                           </div>
                       </div>

2016.8.7

in the morning
</small>

                               </div>
We use semiquantitive RT-PCR

to amplify hhl1 to see how much hhl1 is transcripted into mRNA. 

                                       We failed to get any hhl1.   There may be something wrong with the process of transformation.


                           </div>
                       </div>

2016.8.10

PCR our target DNA fragments:cop1,pif1,phyB,hhl1,pHhl1-hhl1,flu. 

                                       We will check the result tomorrow.


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