Team:WHU-China/Notebook

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<img src="/wiki/images/6/6f/Whu-Protocols.png"/>
<img src="/wiki/images/6/6f/Whu-Protocols.png"/>
<img src="https://static.igem.org/mediawiki/2011/c/c6/Whu-protologo1.png" style="position:absolute;left:20px;top:40px;width:300px;height:360px;">
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<div style="position:absolute;top:60px;right:30px;width:620px;font-size:15px;">
•          Dealing with biobricks</br>
•          Dealing with biobricks</br>
•          Preparation of competent cells (Using Calcium Chloride)</br>
•          Preparation of competent cells (Using Calcium Chloride)</br>
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•          Others............
•          Others............
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<a href="https://2011.igem.org/Team:WHU-China/Notebook/Protocols"><img style="position:absolute;top:380px;right:100px;width:90px;height:30px" src="/wiki/images/1/14/Whu-More.png "></a>
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<a href="https://2011.igem.org/Team:WHU-China/Notebook/Protocols"><img style="position:absolute;top:380px;right:100px;width:80px;height:30px" src="/wiki/images/1/14/Whu-More.png "></a>
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<div id="daytoday">
<div id="daytoday">
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<a href="https://2011.igem.org/Team:WHU-China/Notebook/Progress"><img style="position:absolute;top:380px;right:100px;width:90px;height:30px" src="/wiki/images/1/14/Whu-More.png "></a>
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<img src="/wiki/images/8/83/Whu-Progress.png"/>
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<img src="/wiki/images/0/02/Whu-Progress.jpg" style="position:absolute;left:20px;top:53px;width:400px;height:320px;">
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<div style="position:absolute;top:60px;right:30px;width:520px;font-size:15px;">
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The forth layer assembly and assembled CHL1-3:</br>
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&nbsp;&nbsp;&nbsp;&nbsp;8/26 extract plasmid: 009-13,009-3. Enzyme digest 009-13,009- 3. Result shows that it is not cut completely .PCR 20 + 2 positive control.Transformation: 2 M-13 J-2 M-20 F, PSB, 2 K-16 MIV, CHL1-CHL3 (DH5 α and TrpR defects strains)</br>
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&nbsp;&nbsp;&nbsp;&nbsp;8/27 extract plasmid: 8H, 2M-14 H-2M. enzyme digest 009-3, 8H, 2 M-14 H-2 M. Ligate 009-2-3.Identification of the transformation of PCR</br> &nbsp;&nbsp;&nbsp;&nbsp;8/26 ligation results. Cut CHL1 religate CHL1-CHL3, CHL1-CHL4.Enzyme cut PCB, 23 L, RLS1.Transformation: CHL1-CHL3 (DH5 α and TrpR defects plant), CHL1-CHL4 (DH5 α and TrpR defects plant), 009-2-3, controls.</br>
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&nbsp;&nbsp;&nbsp;&nbsp;8/28 repeat 27th enzymes digest,ligated 009-2-3. Transformation: 18 M-RLS1, 18 O-RLS1, 2 G Ⅱ-RLS1, PSB-23 L, 009-2-3, CHL1-CHL3 (TrpR defects plant), CHL1-CHL4 (TrpR defects strains) controls. Reserve  RLSF-2, PSB-1.....
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<a href="https://2011.igem.org/Team:WHU-China/Notebook/Daytoday"><img style="position:absolute;top:380px;right:100px;width:80px;height:30px" src="/wiki/images/1/14/Whu-More.png "></a>
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<a href="https://2011.igem.org/wiki/index.php?title=Team:WHU-China/Notebook&action=edit"><img class="whucontact" style="position:relative;left:636px;top:38px;z-index:10;" src="/wiki/images/2/2a/Whu-Home_r1_c1.png"/></a>
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<a href="Mailto:gubrian890512@gmail.com"><img class="whucontact" style="position:relative;left:660px;top:38px;z-index:10;" src="/wiki/images/9/97/Whu-Home_r1_c3.png"/></a>
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<a href="https://igem.org/Main_Page"><img class="whucontact" style="position:relative;left:676px;top:38px;z-index:10;" src="/wiki/images/0/03/Whu-Home_r1_c5.png"/></a>
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<img id="bb" src="/wiki/images/3/3c/Whu-build.jpg"/>
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{{WHUcss}}
{{WHUcss}}

Latest revision as of 02:17, 29 October 2011

• Dealing with biobricks
• Preparation of competent cells (Using Calcium Chloride)
• Plasmid Extraction(Using Plasmid Mini Kit)
• Transformation (heat-shock method)
• Cleavage with two restriction enzymes
• Get Extraction
• Identification of DNA by Electrophoresis
• Ligation
• PCR of the colony
• Identification of DNA by Sequencing
• Others............
The forth layer assembly and assembled CHL1-3:
    8/26 extract plasmid: 009-13,009-3. Enzyme digest 009-13,009- 3. Result shows that it is not cut completely .PCR 20 + 2 positive control.Transformation: 2 M-13 J-2 M-20 F, PSB, 2 K-16 MIV, CHL1-CHL3 (DH5 α and TrpR defects strains)
    8/27 extract plasmid: 8H, 2M-14 H-2M. enzyme digest 009-3, 8H, 2 M-14 H-2 M. Ligate 009-2-3.Identification of the transformation of PCR
    8/26 ligation results. Cut CHL1 religate CHL1-CHL3, CHL1-CHL4.Enzyme cut PCB, 23 L, RLS1.Transformation: CHL1-CHL3 (DH5 α and TrpR defects plant), CHL1-CHL4 (DH5 α and TrpR defects plant), 009-2-3, controls.
    8/28 repeat 27th enzymes digest,ligated 009-2-3. Transformation: 18 M-RLS1, 18 O-RLS1, 2 G Ⅱ-RLS1, PSB-23 L, 009-2-3, CHL1-CHL3 (TrpR defects plant), CHL1-CHL4 (TrpR defects strains) controls. Reserve RLSF-2, PSB-1.....
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