Team:USTC-China/Notebook

From 2011.igem.org

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{{Team:USTC-China/temp}}
 
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{{Team:USTC-China/temp2}}
 
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/* end twitter */
 
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/* TEAM PAGE */
 
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.bio {
 
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<html>
 
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<head>
 
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<!-- TODO: add pre/n buttons -->
 
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$(document).ready(function(){
 
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</head>
 
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<body>
 
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<div id="col_nav">
 
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    <div class="col_list">
 
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        <h2>Weeks</h2>
 
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<ul>
 
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    <li id="yhl">Jun 28-July 2</li>
 
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    <li id="swl">July 3-July 9</li>
 
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    <li id="zlc">July 10-July 16</li>
 
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    <li id="fzz">July 17-July 23</li>
 
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    <li id="shl">July 24-July 30</li>
 
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    <li id="ml">July 31-Aug 6</li>
 
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    <li id="mps">Aug 7-Aug 13</li>
 
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            <li id="yfl">Aug 14-Aug 20</li>
 
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    <li id="dz">Aug 21-Aug 27</li>
 
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    <li id="lna">Aug 28-Sep 3</li>
 
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    <li id="jhp">Sep 4-Sep 10</li>
 
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    <li id="xlz">Sep 11-Sep 17</li>
 
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            <li id="wyy">Sep 18-Sep 24</il>
 
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    <li id="ytl">Sep 25-Oct 1</li>
 
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            <li id="nw">Oct 2-Oct 8</li>
 
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            <li id="yz">Oct 9-Oct 15</il>
 
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</ul>
 
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    </div><!-- end undergrads -->
 
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</div>
 
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<div class="bio" id="yhlbio">
 
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<h1>Jun 28-July 2</h1>
 
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2011.6.28
 
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----
 
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cultivate the bacteria of [https://2011.igem.org//Team:USTC-China/cheZ cheZ] deficiency(RP1616),and the Control group(RP437).
 
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check the resistibility of RP1616 and RP437
 
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result:both of the two groups are of none resistibility.
 
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members:Siwei Luo, Fangzhou Zhao, Zhilin Chen...
 
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2011.6.29
 
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----
 
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check the motility of RP1616 strain and RP437 strain
 
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cultivate Top10 strain
 
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result: the bacterial plaque of RP1616 is smaller than that of RP437, implying that RP1616 is of [https://2011.igem.org//Team:USTC-China/cheZ cheZ] deficiency, causing the decreased motility of the bacteria.
 
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6.30
 
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----
 
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prepare for the competent cell of Top10 strain cultivated on 6.29
 
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extract the genome of Top10 strain and use it as complates to run PCR of CheZ
 
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result: the concentration of PCR result is too low to continue the experiments.
 
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7.1
 
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----
 
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conduct PCR of CheZ again
 
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ligate the CheZ DNA segment to plasmid PSb1C3, and transform it into Top10 competent cells
 
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result:as the picture shows.
 
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7.2
 
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----
 
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pick up single colony from the Petri dish to reproduce, in which the competent cells are cultivated and use them colony PCR
 
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result:The bacteria grow up well on dishes with chloromycetin resistency,verifying that the transformation is successfull
 
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while the concentration of PCR product is pretty high,demonstrating that the plasmids indeed contains the sagment CheZ.
 
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<div class="bio" id="swlbio">
 
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<h1>July 3-July 9</h1>
 
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</html>
 
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7.3
 
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----
 
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extract the plasmids from the reproduced bacteria and send it to check the base sequence
 
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result: the concentration of the plasmids extracted is about 500ng/uL
 
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7.4
 
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extract the standard parts LuxPR and Terminator from plate sent by iGEM registry 2010 and put them into Top10 strain by transformation
 
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(LuxPr: Plate 2,24C  Terminator : Plate 1,6O)
 
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result: the bacteria on the plate with Amp resistency grows well
 
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7.5
 
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----
 
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pick up single colony from the Petri dish to reproduce in the liquid culture medium
 
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7.6
 
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extract plasmids containing LuxPR and Terminator and make enzyme digestion with EcoR1 and Spe1 in order to validate the existence of the two parts
 
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recieve part Toggle Switch and rbs-Ci-ter from PKU and conduct transformation
 
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result: Due to the short length of LuxPR and Terminator, the result of the enzyme digestion is hard to examine by electrophoresis.
 
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7.7
 
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----
 
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result: there are a few colonies with part Toggle switch grown while no colony of rbs-Ci-ter appeared.
 
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7.8
 
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----
 
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pick up single colony with part Toggle switch .
 
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extract the standard parts LuxPR and Terminator from plate sent by iGEM registry 2011 and put them into Top10 strain by transformation
 
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7.9
 
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----
 
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pick up single colony with LuxPR & terminator.
 
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extract plasmids containing toggle switch to conduct PCR.
 
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process PCR with plasmids aptamer-CheZ
 
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result:as the picture shows.
 
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the base sequence of CheZ extracted before is proved right.
 
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</div>
 
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<div class="bio" id="zlcbio">
 
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<h1>July 10-July 16</h1>
 
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</html>
 
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7.10
 
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ligate aptamer-CheZ with PSB1C3 plasmids
 
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extract plasmids containing LuxPR and terminator
 
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result:the concentractions of plasmids LuxPR and terminator are both 600ng/ul
 
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7.11
 
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----
 
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transform PSB1C3 containing aptamer-CheZ into bacteria and cultivate.
 
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7.12
 
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carry out Double digestion of toggle switch and pSB1c3 and ligate them together.
 
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pick up single colony from bacteria cultivated yesterday.
 
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result: Both the location and the brightness of the electrophoretic bands correspond with our expectation.
 
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7.14
 
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----
 
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transform the part rbs-ci-ter sent from PKU again into bacteria.
 
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check the colors of the colonies containing toggle switch.
 
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result: the ratio of red vs green is 8:25, verifying the function of toggle switch.
 
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7.15
 
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----
 
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pick up the single colony from the plate yesterday and ten hours later extract the plasmids containing Ci-terminator.
 
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7.16
 
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----
 
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conduct PCR with the plasmids yesterday to reproduce rbs-ci-Term.
 
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result:failure
 
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<html>
 
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</div>
 
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<div class="bio" id="fzzbio">
 
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<h1>July 17-July 23</h1>
 
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7.18-7.21
 
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----
 
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ligate the standard part Terminator to the end of toggle switch
 
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result: we conduct a enzyme digestion with EcoR1 and Pst1 to the final ligation product and the electrophoresis result shows that the ligation is successful.
 
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<div class="clear"></div>
 
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</div>
 
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<div class="bio" id="shlbio">
 
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<h1>July 24-July 30</h1>
 
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7.22-7.29
 
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----
 
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perform the following experiments: PCR of rbs-ci-ter, enzyme digestion(E,S),ligation with LuxPR(standard part from IGEM registry digested by E,X),tranformation,plasmids extraction.
 
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result: the concentration of plasmids with ligation gene is as high as ...
 
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<div class="clear"></div>
 
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</div>
 
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<div class="bio" id="mlbio">
 
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<h1>July 31-Aug 6</h1>
 
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7.30-8.3
 
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----
 
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PCR of LuxPR-rbs-ci-ter, enzyme digestion(E,S),ligation with LuxPR(standard part from IGEM registry digested by E,X),tranformation,plasmids extraction.
 
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</div>
 
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<div class="bio" id="mpsbio">
 
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<h1>Aug 7-Aug 13</h1>
 
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<div class="bio" id="yflbio">
 
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<h1>Aug 14-Aug 20</h1>
 
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</div>
 
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<div class="bio" id="nwbio">
 
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<h1></h1>
 
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Latest revision as of 18:06, 5 October 2011