Team:OUC-China/Lab note

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       <p style="font-weight:normal;"><font size="3px">1. Culture the E.coli cells: Psb1c3.<br/>
       <p style="font-weight:normal;"><font size="3px">1. Culture the E.coli cells: Psb1c3.<br/>
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</section>
 
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<section id="Week5">
 
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  <div class="page-header">
 
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            <h1>Week5</h1>
 
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          </div>
 
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  <div class="row">
 
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  <div class="span9">
 
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    <div class="row">
 
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      <h4>July 21st</h4>
 
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      <div class="span9"><p style="font-weight:normal;"><font size="3px">Person: Wenjun Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Agarose electrophoresis: detect the plasmid PCR product.<br/>
 
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2. PCR: Get mamB sequence from PCR product.<br/>
 
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3. PCR: Get mamQ sequence from PCR product.<br/>
 
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4. PCR: Get mamJ sequence from PCR product to finish the Over-Lap PCR. <br/>
 
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5. Agarose electrophoresis: detect the plasmid PCR product.<br/>
 
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6. Product purification: purify the PCR product.
 
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   </font></p></div>
 
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  <div class="span9"><p style="font-weight:normal;"><font size="3px">Person: Yu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Bacterination: E.coli with part K1059003-B0035-GFPlva-K1059004-B0015 pSB1AK3 as backbone; E.coli with RFP, pSB1C3 as backbone.
 
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   </font></p></div>
 
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    <div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Plasmid miniprep: plasmid Extraction of PSB1C3 for 5 tubes.<br/>2. Electrophoresis: gel is good.<br/>3. Equip the medium: Equip the medium of LB (solid).
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. PCR production purification: 16SrRNA from AMB-1.<br/>2. SDS page gel electrophoresis: the purified PCR production.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xiaodong Zhong
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. PCR genome for mamI gene & PCR purification.<br/>2. PCR genome for mamB gene.
 
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   </font></p></div>
 
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    <h4>July 22nd</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xue Sun
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Plasmid miniprep: Plasmid pSB1AK3 with K1059003-B0035-GFPlva-K1059004-B0015, and pSB1C3 with RFP.<br/>2. Enzyme digestion.<br/>3. Gel extraction.<br/>4. Recombination: Get part K1059003-B0035-GFPlva-K1059004-B0015, pSb1C3 as backbone.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1.Enzyme digestion: enzyme digestion of PSB1C3 for 25ul system.<br/>2.Gel recovery: gel recovery of the product of enzyme digestion.<br/>3.Bacterination: bacterinate PSB1C3-RFP, RNA1 into tubes of 10ml.<br/>4.Pick bacteria: pick bacteria of PSB1C3-RFP for conservation.<br/>5.Genomic DNA extraction: extracting genomic DNA of E.coli.<br/>6. Electrophoresis: gel is bad.<br/>7. Bacterination: bacterinate PSB1C3-RFP, RNA1 into tubes of 10ml for 4 tubes.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Ligation: ligate the purified PCR production with T vector.<br/>2. Transformation of Top10.
 
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   </font></p></div>
 
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  <h4>July 23rd</h4>
 
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  <div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Extraction of genomic DNA from E.coli.<br/>2. Pick the bacteria and culture the E.coli cells for four hours.<br/>3. PCR: the cultured E.coli cells.<br/>4. SDS page gel electrophoresis: PCR production.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Plasmid miniprep: plasmid Extraction of PSB1C3 -RFP for 5 tubes.<br/>
 
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2. Enzyme digestion: enzyme digestion of PSB1C3 for 50ul system of 3 tubes, use EcoR1, Pst1.3. Electrophoresis: gel is good.<br/>4. Genomic DNA extraction: extracting genomic DNA of E.coli.<br/>5. Electrophoresis: gel is bad.<br/>6. Gel recovery: gel recovery of PSB1C3.<br/>7. Electrophoresis: gel is good for 50ng/ul each.<br/>8. Bacterination: bacterination of E.coli for 6 tubes.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qiu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Transformation: Plate cultivation for 14h.
 
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   </font></p></div>
 
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  <h4>July 24th</h4>
 
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  <div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Extraction of genomic DNA from E.coli.<br/>2. PCR: the cultured E.coli cells.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Genomic DNA extraction: extracting genomic DNA of E.coli.<br/>2. Electrophoresis: gel is bad.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Wenjun Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Agarose electrophoresis: detect the plasmid PCR product.<br/>2. PCR: Get mamI sequence from PCR product.<br/>3. PCR: Get mamL sequence from PCR product.<br/>4. PCR: Get mamK sequence from PCR product to finish the Over-Lap PCR. <br/>5. Agarose electrophoresis: detect the plasmid PCR product.<br/>6. Product purification: purify the PCR product.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Yu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Pick bacteria and PCR examining, the aim fragment is 1409bp.<br/>2. Sequencing.
 
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   </font></p></div>
 
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  <h4>July 25th</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Wenjun Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. PCR: Try to get mamAB gene cluster sequence from AMB-1 bacterial strain.<br/>2. Get the genome of AMB-1 with TaKaRa MINI Best bacteria genome DNA Extraction Ver2.0.<br/>3. PCR: Try to get mamAB gene cluster sequence from AMB-1 genome.<br/>4. Agarose electrophoresis: detect the plasmid PCR product.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xue Sun
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. d1 to d6 as primer PCR, get part K1059005.<br/>2. Purification.<br/>3. Recombine PCR product to pMD-19 T vector.<br/>4. Transformation.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1.Equip the medium:Equip the medium of AMB-1.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. SDS page gel electrophoresis: PCR production and the genomic DNA.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xiaodong Zhong
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Transformation of E.coli (top10): <br/>
 
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Part: mamK+T vector <br/>
 
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Part: mamI+T vector<br/>
 
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Part: mamL+T vector<br/>
 
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Part: mamQ+T vector<br/>
 
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Part: mamB+T vector (failure)
 
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   </font></p></div>
 
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<h4>July 26th</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Yu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Blue-white selection.<br/>2. PCR examining, the aim fragment is 388bp.<br/>3. Sequencing.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Bacterination: bacterinate PSB1C3-RFP into tubes of 10ml.
 
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   </font></p></div>
 
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<h4>July 27th</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qiu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Measuring Growth curve, E.coli with RFP, pSB1C3 as plasmid backbone.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Extraction of genomic DNA from E.coli.<br/>2. SDS page gel electrophoresis: the genomic DNA.
 
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   </font></p></div>
 
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  </div>
 
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  </div>
 
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  </div>
 
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<br /><br /></br>
 
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</section>
 
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  <section id="Week6">
 
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  <div class="page-header">
 
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            <h1>Week6</h1>
 
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          </div>
 
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  <div class="row">
 
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  <div class="span9">
 
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    <div class="row">
 
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      <h4>July 28th</h4>
 
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      <div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qiu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Measure Growth curve.
 
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   </font></p></div>
 
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  <div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Bacterination: bacterinate PSB1C3-RFP, E.coil with single-1 protection sequence into tubes of 10ml.
 
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   </font></p></div>
 
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    <div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Culture the E.coil cells: PSB1C3.
 
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   </font></p></div>
 
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<h4>July 29th</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Yu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Bacterial culturing.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Wenjun Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. PCR: Get mamB sequence from AMB-1 bacterial strain.<br/>2. PCR: Get mamJ sequence from AMB-1 bacterial strain.<br/>3. PCR: Get mamE sequence from AMB-1 bacterial strain.<br/>4. PCR: Get mamE sequence from AMB-1 bacterial strain.<br/>5. Agarose electrophoresis: detect the DNA digestion product.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Plasmid miniprep: plasmid Extraction of PSB1C3-RFP, E.coil with single-1 protection sequence for 2 tubes each.<br/>2. Enzyme digestion: enzyme digestion of PSB1C3 and E.coil with single-1 protection sequence for 25ul system of 2 tubes each, use EcoR1, Pst1.<br/>3. Electrophoresis: gel is good.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Miniprep: Psb1c3.<br/>2. Digestion: digest the plasmid with EcoRI and Pst-HF.<br/>3. SDS page gel electrophoresis.<br/>4. Gel extraction of DNA.
 
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   </font></p></div>
 
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<h4>July 30th</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qiu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Lstwenmin1 to Lstwenmin4 as primer PCR, get part K1059006.<br/>2. Purification.<br/>3. Recombine PCR product to pMD-19 T vector.<br/>4. Transformation.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Transform: transform the ligastion (single-1 protection sequence-PSB1C3) product into Top 10.<br/>2. Bacterial coating.
 
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   </font></p></div>
 
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<h4>July 31st</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xue Sun
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Blue-white selection.<br/>2. PCR and electrophoresis examining, the aim fragment is 388bp and 210.<br/>3. Sequencing.<br/>4. The PCR result a fragment at 120bp, so we failed the part k1059006. We designed another two primers and be ready for more PCR.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Kaili Qin
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Pick bacteria: pick bacteria for PCR.<br/>2. PCR: use PCR to detect the bacteria we want (single-1 protection sequence-PSB1C3)), 25ul system.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Glycerol stocks: Psb1c3, 2 tubes.<br/>2. Miniprep: Psb1c3.<br/>3. Digestion: digest the plasmid with EcoRI and Pst-HF.<br/>4. Gel extraction of DNA.
 
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   </font></p></div>
 
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<h4>August 1st</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Yu Wang
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Bacterination.<br/>2. E.coil with part K1059005 PMD-19 T vector as backbone.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xiaodong Zhong
 
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   </font></p>
 
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      <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. PCR genome for mamB, mamJ, mamE, mamC gene.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Wenjun Wang
 
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   </font></p>
 
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    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. PCR: Get mamC sequence from AMB-1 bacterial strain.<br/>2. Agarose electrophoresis: detect the mamC PCR product.<br/>3. Product purification: purify the PCR product.<br/>4. PCR: Get the mamC sequence which is more nicety from the product of product purification.
 
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   </font></p></div>
 
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<h4>August 2nd</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xue Sun
 
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   </font></p>
 
-
    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Plasmid minprep.<br/>2. Enzyme digestion.<br/>3. Recombination: Get part K1059005-B0035-GFPlva-K1059004-B0015, pSB1C3 as backbone.
 
-
   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xiaodong Zhong
 
-
   </font></p>
 
-
    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Transformation of E.coil (top10): mamB+T vector.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Wenjun Wang
 
-
   </font></p>
 
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    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. PCR:Get mamB, mamE sequence from AMB-1 bacterial strain.<br/>2. Get the genome of AMB-1.<br/>3. Transform: transform the ligastion product into Top 10.
 
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   </font></p></div>
 
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<h4>August 3rd</h4>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qiu Wang
 
-
   </font></p>
 
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    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Transformation and Bacterination.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Qianyun Lu
 
-
   </font></p>
 
-
    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Miniprep: Psb1c3, 6 tubes.<br/>2. Digestion: digest the plasmid with EcoRI and Pst-HF, XbaI and Pst-HF.<br/>3. SDS page gel electrophoresis.<br/>4. Gel extraction of DNA.
 
-
   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Xiaodong Zhong
 
-
   </font></p>
 
-
    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. E. coli Cell Culture: Top10 (mamB+T vector) Resistance: Ampicillin Medium: LB.<br/>2. mamK PCR purification: Get mamK gene.<br/>3. PCR genome for mamB: 16x50ul PCR system. Get mamB gene.
 
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   </font></p></div>
 
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<div class="span9"><p style="font-weight:normal;"><font size="3px">Person:Wenjun Wang
 
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   </font></p>
 
-
    <p style="font-weight:normal;"><font size="3px">Experiment:<br/>1. Agarose electrophoresis: detect the PCR product.<br/>2. Get the genome of AMB-1.<br/>3. Agarose electrophoresis: detect the genome product.<br/>4. PCR: to detect the product of transform.<br/>5. PCR: Get mamC sequence from AMB-1 bacterial strain.<br/>6. PCR: Get 16srDNA sequence from AMB-1 bacterial strain to detect the strain.<br/>7. Agarose electrophoresis: detect the genome product.
 
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   </font></p></div>
 
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  </div>
  </div>
  </div>
  </div>

Revision as of 08:05, 26 September 2013

 










 
 
Lab-note






June 26th

Person: Wenjun Wang

 

Experiment:
1. Get the AMB-1 bacteria strain from Nanjing University.
2. Keep the strain in 30℃.




June 30th

Person: Wengjun Wang    

Experiment:
1. PCR: Get 16srDNA sequence from AMB-1 bacteria genome to detect the strain.
2. Agarose electrophoresis: detect the PCR product.
3. Transform: transform the ligastion product into Top 10 to prepare for sequence.    




July 13th ~ 15th

Person: Wenjun Wang

 

Experiment:
1. Get the AMB-1 bacteria strain from Nanjing University.
2. Keep the strain in 30℃.




June 30th

Person: Kaili Qin    

Experiment:
  1. Equip the medium: Equip the medium of Magnetospirillum Magneticum AMB-1.
   

July 20th

Person: Wenjun Wang    

Experiment:
1. PCR: Get mamJ sequence from PCR product to finish the Over-Lap PCR.
2. Agarose electrophoresis: detect the plasmid PCR product.
3. Transform: transform the ligastion product into Top 10 to prepare for sequence.
4. PCR: Get mamK sequence from PCR product to finish the Over-Lap PCR.
5. Agarose electrophoresis: detect the PCR product.
6. Product purification: purify the PCR product.
7. PCR: Get mamAB sequence from AMB-1 bacteria genome.    

Person: Qianyun Lu    

1. Culture the E.coli cells: Psb1c3.