Team:USTC-China/groupmeetings

From 2012.igem.org

(Difference between revisions)
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         <h2>Weeks</h2>
         <h2>Weeks</h2>
<ul>
<ul>
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    <li id="yhl">Jun 28-July 2</li>
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    <li id="yhl">18th,Feb</li>
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    <li id="swl">July 3-July 9</li>
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    <li id="swl">25th,Feb</li>
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    <li id="zlc">July 10-July 16</li>
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    <li id="zlc">3rd, Mar</li>
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    <li id="fzz">July 17-July 23</li>
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    <li id="fzz">10th,Mar</li>
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    <li id="shl">July 24-July 30</li>
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    <li id="shl">17th,Mar</li>
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    <li id="ml">July 31-Aug 6</li>
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    <li id="ml">24th,Mar</li>
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    <li id="mps">Aug 7-Aug 13</li>
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    <li id="mps">7th, Apr</li>
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             <li id="yfl">Aug 14-Aug 20</li>
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             <li id="yfl">14th,Apr</li>
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    <li id="dz">Aug 21-Aug 27</li>
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    <li id="dz">6th, May</li>
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    <li id="lna">Aug 28-Sep 3</li>
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    <li id="lna">13th,May</li>
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    <li id="jhp">Sep 4-Sep 10</li>
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    <li id="jhp">21st,May</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>
</ul>
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<div class="bio" id="yhlbio">
<div class="bio" id="yhlbio">
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<h1>Jun 28-July 2</h1>
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<h1>18th, Feb</h1>
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<p>2011.6.28
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<p>Place: Room 363, Life Science building
</p>
</p>
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<hr />
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<p>Instructor: Mr.Hong
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<p>cultivate the bacteria of <a href="http://partsregistry.org/Part:BBa_K557000" class="external text" rel="nofollow">cheZ</a> 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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</p>
</p>
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<p>2011.6.29</p>
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<p>Recordist: Wuyang Chen
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<hr />
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<p>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 <a href="http://partsregistry.org/Part:BBa_K557000" class="external text" rel="nofollow">cheZ</a> deficiency, causing the decreased motility of the bacteria.</p>
+
-
<p><br />
+
-
6.30
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</p>
</p>
<hr />
<hr />
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<p>prepare for the competent cell of Top10 strain cultivated on 6.29
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<p>I. Discuss about our project:
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extract the genome of Top10 strain and use it as complates to run PCR of CheZ
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1. SNS:<br>
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result: the concentration of PCR result is too low to continue the experiments.</p>
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To complete this project is hardly possible. While it is likely to realize part of it, such as a system which is able to create and sense signals. But we need to confer our project some new meanings.
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<p>7.1</p>
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1) About making a signal disappear, according to Mr. Hong, we can add a rapid degradation label onto the signal.
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<hr />
+
2) We are afraid that the colony is too small to create an appropriate concentration gradient. According to Mr. Hong, it won’t be a big problem.
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<p>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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2. Solve the graph colouring problem using E.coli
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result:as the picture shows.
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1) In fact, we will only use E.coli to emit light in different colors, and all the algorithm for solving graph coloring problem is processed artificially.
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</p><p>7.2
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2) We need to consult professors in Chemistry Department about constructing structures using ssDNA.
 +
3) The scale of DNA strand is far too small than that of E.coli.
 +
4) The fusion protein of Zn finger and the Ag43 would be very useful. We can think about how to make use of it.
 +
 
 +
3. Fatigue of specific stimulus
 +
1) comK is a protein produced by bacillus subtilis and “competence” is a state of it. The possibility of turning into competence state is proportional to the amount of comK. We can substitute the protein involved in competence state for the protein we need.
 +
2) The system can be used as a special lock.
 +
3) How to make circuit one recover its function as quick as possible?
 +
a) by means of protease?
 +
 
 +
II. Set up groups to continue brainstorm
 +
 
 +
III. Make plans for the whole term.
</p>
</p>
-
<hr />
+
 
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<p>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.</p>
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-
<p><br />
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<div class="clear"></div>
<div class="clear"></div>

Revision as of 08:02, 20 September 2012

GROUP MEETINGS

Weeks

  • 18th,Feb
  • 25th,Feb
  • 3rd, Mar
  • 10th,Mar
  • 17th,Mar
  • 24th,Mar
  • 7th, Apr
  • 14th,Apr
  • 6th, May
  • 13th,May
  • 21st,May

18th, Feb

Place: Room 363, Life Science building

Instructor: Mr.Hong

Recordist: Wuyang Chen

I. Discuss about our project: 1. SNS:
To complete this project is hardly possible. While it is likely to realize part of it, such as a system which is able to create and sense signals. But we need to confer our project some new meanings. 1) About making a signal disappear, according to Mr. Hong, we can add a rapid degradation label onto the signal. 2) We are afraid that the colony is too small to create an appropriate concentration gradient. According to Mr. Hong, it won’t be a big problem. 2. Solve the graph colouring problem using E.coli 1) In fact, we will only use E.coli to emit light in different colors, and all the algorithm for solving graph coloring problem is processed artificially. 2) We need to consult professors in Chemistry Department about constructing structures using ssDNA. 3) The scale of DNA strand is far too small than that of E.coli. 4) The fusion protein of Zn finger and the Ag43 would be very useful. We can think about how to make use of it. 3. Fatigue of specific stimulus 1) comK is a protein produced by bacillus subtilis and “competence” is a state of it. The possibility of turning into competence state is proportional to the amount of comK. We can substitute the protein involved in competence state for the protein we need. 2) The system can be used as a special lock. 3) How to make circuit one recover its function as quick as possible? a) by means of protease? II. Set up groups to continue brainstorm III. Make plans for the whole term.

July 3-July 9

7.3

extract the plasmids from the reproduced bacteria and send it to check the base sequence result: the concentration of the plasmids extracted is about 500ng/uL

7.4

extract the standard parts LuxPR and Terminator from plate sent by iGEM registry 2010 and put them into Top10 strain by transformation (LuxPr: Plate 2,24C Terminator : Plate 1,6O) result: the bacteria on the plate with Amp resistency grows well 7.5

pick up single colony from the Petri dish to reproduce in the liquid culture medium


7.6

extract plasmids containing LuxPR and Terminator and make enzyme digestion with EcoR1 and Spe1 in order to validate the existence of the two parts recieve part Toggle Switch and rbs-Ci-ter from PKU and conduct transformation

result: Due to the short length of LuxPR and Terminator, the result of the enzyme digestion is hard to examine by electrophoresis.

7.7

result: there are a few colonies with part Toggle switch grown while no colony of rbs-Ci-ter appeared.

7.8

pick up single colony with part Toggle switch . extract the standard parts LuxPR and Terminator from plate sent by iGEM registry 2011 and put them into Top10 strain by transformation

7.9

pick up single colony with LuxPR & terminator. extract plasmids containing toggle switch to conduct PCR.

process PCR with plasmids aptamer-CheZ result:as the picture shows. the base sequence of CheZ extracted before is proved right.


July 10-July 16

7.10

ligate aptamer-CheZ with PSB1C3 plasmids

extract plasmids containing LuxPR and terminator result:the concentractions of plasmids LuxPR and terminator are both 600ng/ul

7.11

transform PSB1C3 containing aptamer-CheZ into bacteria and cultivate. 7.12

carry out Double digestion of toggle switch and pSB1c3 and ligate them together.

pick up single colony from bacteria cultivated yesterday.

result: Both the location and the brightness of the electrophoretic bands correspond with our expectation.

7.14

transform the part rbs-ci-ter sent from PKU again into bacteria. check the colors of the colonies containing toggle switch. result: the ratio of red vs green is 8:25, verifying the function of toggle switch.

7.15

pick up the single colony from the plate yesterday and ten hours later extract the plasmids containing Ci-terminator.


7.16

conduct PCR with the plasmids yesterday to reproduce rbs-ci-Term. result:failure


July 17-July 23

7.18-7.21 ---- ligate the standard part Terminator to the end of toggle switch 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.

July 24-July 30

7.22-7.29 ---- 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. result: the concentration of plasmids with ligation gene is as high as ...

July 31-Aug 6

7.30-8.3 ---- 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.

Aug 7-Aug 13

Aug 14-Aug 20

backtotop

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