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

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<body>
 
<body>
 
     <div class="wrapper wrapper-content">
 
     <div class="wrapper wrapper-content">
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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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</div> -->
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<div class="navbar-collapse collapse" id="navbar">
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<ul class="nav navbar-nav navbar-right">
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<li class="dropdown">
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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>
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<li class="dropdown">
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<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> TEAM <span class="caret"></span></a>
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<ul role="menu" class="dropdown-menu">
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<li><a href="#">Team members</a></li>
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</ul>
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<li class="dropdown">
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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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<ul role="menu" class="dropdown-menu">
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<li><a href="#">Project description</a></li>
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<li><a href="#">制取拟南芥DNA</a></li>
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<li><a href="#">酶切</a></li>
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<li><a href="#">制取拟南芥原生质体</a></li>
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<li><a href="#">result</a></li>
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<li><a href="#">跑胶图片</a></li>
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</ul>
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<li class="dropdown">
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<a aria-expanded="false" role="button" href="#" class="dropdown-toggle" data-toggle="dropdown"> PARTS <span class="caret"></span></a>
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<li><a href="#">Part1:PHhl1-hhl1-flu</a></li>
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<li><a href="#">Part2:pPhyB-hhl1-flu</a></li>
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<li><a href="#">Part3:pPif1-hhl1-flu</a></li>
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<li><a href="#">Part4:pCop1-hhl1-flu</a></li>
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</ul>
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<li><a href="GDSYZX-United/NOTEBOOK/day_note">1.Day notes</a></li>
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<li><a href="#">实验过程记录</a></li>
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<li><a href="#">3.Protocol</a></li>
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</ul>
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<li><a href="#">wiki相册</a></li>
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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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</ul>
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</ul>
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Revision as of 07:11, 23 September 2016

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.