Difference between revisions of "Team:TJU China/Notebook/Group1"

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         .notebook_month p {
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         .notebook_month_p {
 
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         }
 
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         .notebook_week p {
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         .notebook_week_p{
 
             text-align: left;
 
             text-align: left;
 
             font-size: 20px;
 
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     </div>
 
     </div>
 
     <div style=" margin-top:28px;z-index:10; border-top: solid #4e72b8 2px;width: 100%; position: fixed;"></div>
 
     <div style=" margin-top:28px;z-index:10; border-top: solid #4e72b8 2px;width: 100%; position: fixed;"></div>
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     <div class="notebook_month">
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     <div class="notebook_month"> class="notebook_month_p"
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         <p>March</p>
+
         <div class="notebook_month_p">March</div>
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         <p>April</p>
+
         <div class="notebook_month_p">April</div>
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         <p>May</p>
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         <div class="notebook_month_p">May</div>
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         <p>June</p>
+
         <div class="notebook_month_p">June</div>
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         <p>July</p>
+
         <div class="notebook_month_p">July</div>
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         <p>August</p>
+
         <div class="notebook_month_p">August</div>
−
         <p>September</p>
+
         <div class="notebook_month_p">September</div>
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         <p>October</p>
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         <div class="notebook_month_p">October</div>
 
     </div>
 
     </div>
 
     <div class="notebook_timeline">
 
     <div class="notebook_timeline">
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         <button class="notebook_button8" id="notebook_btn8"></button>
 
         <button class="notebook_button8" id="notebook_btn8"></button>
 
     </div>
 
     </div>
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    class="notebook_week_p"
 
     <div class="notebook_information">
 
     <div class="notebook_information">
 
         <div id="notebook_March">
 
         <div id="notebook_March">
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                 </div>
 
                 </div>
 
                 <div id="3notebook_week1" class="notebook_information_word1" style="display:none;">
 
                 <div id="3notebook_week1" class="notebook_information_word1" style="display:none;">
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                     <p>After a lot of document viewing and brainstorms, we finally came up with a clear idea about how we were
+
                     <div class="notebook_week_p">After a lot of document viewing and brainstorms, we finally came up with a clear idea about how we were
 
                         going to carry out our part of project. We decided to construct the RNPs (complex of sgRNA and Cas9
 
                         going to carry out our part of project. We decided to construct the RNPs (complex of sgRNA and Cas9
 
                         protein) and to wrap them with delivery vector in order to improve the gene-editing efficiency of
 
                         protein) and to wrap them with delivery vector in order to improve the gene-editing efficiency of
−
                         the system into the cells.</p>
+
                         the system into the cells.</div>
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="3notebook_week2" class="notebook_information_word2" style="display:none;">
 
                 <div id="3notebook_week2" class="notebook_information_word2" style="display:none;">
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                     <p>After daily meetings and discussions, we exchanged knowledge and ideas from the literatures we read.
+
                     <div class="notebook_week_p">After daily meetings and discussions, we exchanged knowledge and ideas from the literatures we read.
 
                         Comparing different transportation methods, including physical electroporation, chemical nanoparticles,
 
                         Comparing different transportation methods, including physical electroporation, chemical nanoparticles,
 
                         and even virus methods, we decided to improve the transport efficiency of the system by using two-dimensional
 
                         and even virus methods, we decided to improve the transport efficiency of the system by using two-dimensional
−
                         nanomaterials to encapsulate the cas9/sgRNA complex assembled in vitro to transport into the cells.</p>
+
                         nanomaterials to encapsulate the cas9/sgRNA complex assembled in vitro to transport into the cells.</div>
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="3notebook_week3" class="notebook_information_word3" style="display:none;">
 
                 <div id="3notebook_week3" class="notebook_information_word3" style="display:none;">
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                     <p>We began to delve into more detailed and specific knowledge, such as the structure of cas9 protein and
+
                     <div class="notebook_week_p">We began to delve into more detailed and specific knowledge, such as the structure of cas9 protein and
 
                         how it works, the structure and composition of sgRNA and how it is assembled with cas9 protein, the
 
                         how it works, the structure and composition of sgRNA and how it is assembled with cas9 protein, the
−
                         method of primer design, learning how to design sgRNA, and so on.</p>
+
                         method of primer design, learning how to design sgRNA, and so on.</div>
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="3notebook_week4" class="notebook_information_word4" style="display:none;">
 
                 <div id="3notebook_week4" class="notebook_information_word4" style="display:none;">
−
                     <p>After confirming the type of cas9 protein, we started buying and asking other labs if they had the same
+
                     <div class="notebook_week_p">After confirming the type of cas9 protein, we started buying and asking other labs if they had the same
 
                         cas9 protein. While waiting for an answer, we are continuing to refine the questions left over from
 
                         cas9 protein. While waiting for an answer, we are continuing to refine the questions left over from
 
                         last week's knowledge and begin to learn the experimental procedures and necessary knowledge related
 
                         last week's knowledge and begin to learn the experimental procedures and necessary knowledge related
−
                         to protein expression purification.</p>
+
                         to protein expression purification.</div>
 
                 </div>
 
                 </div>
  
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                 </div>
 
                 </div>
 
                 <div id="4notebook_week1" class="notebook_information_word1" style="display:none;">
 
                 <div id="4notebook_week1" class="notebook_information_word1" style="display:none;">
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                     <p>We continued to search for related documents in order to choose the appropriate target sequences of gene
+
                     <div class="notebook_week_p"> We continued to search for related documents in order to choose the appropriate target sequences of gene
 
                         for our project. After the final decision of making eGFP as our interested sequence of gene, we immediately
 
                         for our project. After the final decision of making eGFP as our interested sequence of gene, we immediately
 
                         sent out the primers of sgRNA we designed to GENEWIZ, and we also sent lots of emails to different
 
                         sent out the primers of sgRNA we designed to GENEWIZ, and we also sent lots of emails to different
−
                         labs to see if they can offer us a stable cell strain expressing eGFP protein.</p>
+
                         labs to see if they can offer us a stable cell strain expressing eGFP protein.</div>
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="4notebook_week2" class="notebook_information_word2" style="display:none;">
 
                 <div id="4notebook_week2" class="notebook_information_word2" style="display:none;">
−
                     <p>Meanwhile, we successfully transformed the plasmid pET-NLS-Cas9-6xHis (purchased from Addgene) into E.coli
+
                     <div class="notebook_week_p">Meanwhile, we successfully transformed the plasmid pET-NLS-Cas9-6xHis (purchased from Addgene) into E.coli
 
                         (BL21), then we tried to express and purify the cas9 protein according to the protocol[1]. But the
 
                         (BL21), then we tried to express and purify the cas9 protein according to the protocol[1]. But the
 
                         results were not so ideal.
 
                         results were not so ideal.
 
                         <br> 【1】Ha J S, Lee J S, Jeong J, et al. Poly-sgRNA/siRNA ribonucleoprotein nanoparticles for targeted
 
                         <br> 【1】Ha J S, Lee J S, Jeong J, et al. Poly-sgRNA/siRNA ribonucleoprotein nanoparticles for targeted
−
                         gene disruption[J]. Journal of Controlled Release, 2017, 250:27-35.</p>
+
                         gene disruption[J]. Journal of Controlled Release, 2017, 250:27-35.</div>
 
                 </div>
 
                 </div>
  
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                 </div>
 
                 </div>
 
                 <div id="5notebook_week1" class="notebook_information_word1" style="display:none;">
 
                 <div id="5notebook_week1" class="notebook_information_word1" style="display:none;">
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                     <p>PCR and Gel Purification (TIANgel Midi Purification Kit) of the template of sgRNA, we tried different
+
                     <div class="notebook_week_p">PCR and Gel Purification (TIANgel Midi Purification Kit) of the template of sgRNA, we tried different
−
                         Tm temperatures.</p>
+
                         Tm temperatures.</div>
 
                     <img src="./img/group3May1pic1.PNG">
 
                     <img src="./img/group3May1pic1.PNG">
 
                     <img src="./img/group3May1pic2.PNG">
 
                     <img src="./img/group3May1pic2.PNG">
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                 </div>
 
                 </div>
 
                 <div id="5notebook_week2" class="notebook_information_word2" style="display:none;">
 
                 <div id="5notebook_week2" class="notebook_information_word2" style="display:none;">
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                     <p>After two weeks of hard working, we got our first batch of Cas9 protein.
+
                     <div class="notebook_week_p">After two weeks of hard working, we got our first batch of Cas9 protein.
 
                         <br> At the same time, we tried our first transcription and preparation of sgRNA using T7 High Efficiency
 
                         <br> At the same time, we tried our first transcription and preparation of sgRNA using T7 High Efficiency
 
                         Transcription Kit and EasyPure RNA Purification Kit (TransGene Biotech) but failed. We supposed that
 
                         Transcription Kit and EasyPure RNA Purification Kit (TransGene Biotech) but failed. We supposed that
−
                         the template might be polluted by RNase during the process.</p>
+
                         the template might be polluted by RNase during the process.</div>
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="5notebook_week3" class="notebook_information_word3" style="display:none;">
 
                 <div id="5notebook_week3" class="notebook_information_word3" style="display:none;">
−
                     <p>We searched for information and knowledge of experiments dealing with RNA and then retried the PCR and
+
                     <div class="notebook_week_p">We searched for information and knowledge of experiments dealing with RNA and then retried the PCR and
 
                         Gel Purification of the template according strictly to the RNA principles. However, we failed again.
 
                         Gel Purification of the template according strictly to the RNA principles. However, we failed again.
−
                         <br> On the other hand, we managed to get plasmid encoding AcGFP by using TIANprep Mini Plasmid Kit (TIANGEN).</p>
+
                         <br> On the other hand, we managed to get plasmid encoding AcGFP by using TIANprep Mini Plasmid Kit (TIANGEN).</div>
 
                     <img src="./img/group3May3pic1.png">
 
                     <img src="./img/group3May3pic1.png">
 
                 </div>
 
                 </div>
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                 </div>
 
                 </div>
 
                 <div id="5notebook_week4" class="notebook_information_word4" style="display:none;">
 
                 <div id="5notebook_week4" class="notebook_information_word4" style="display:none;">
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                     <p>We confirmed the best Tm temperature of PCR was 67.2℃ after several preliminary experiments. And this
+
                     <div class="notebook_week_p">We confirmed the best Tm temperature of PCR was 67.2℃ after several preliminary experiments. And this
 
                         time, there were lines shown on the RNA gel, but they were not clear. And the RNA marker (TAKARA)
 
                         time, there were lines shown on the RNA gel, but they were not clear. And the RNA marker (TAKARA)
 
                         didn’t appear exactly as the instruction. Then we moved forward to the In vitro digestion of DNA
 
                         didn’t appear exactly as the instruction. Then we moved forward to the In vitro digestion of DNA
 
                         but failed. Since that the result of the SDS-PAGE of Cas9 protein showed no problem, we considered
 
                         but failed. Since that the result of the SDS-PAGE of Cas9 protein showed no problem, we considered
−
                         that the sgRNA might degraded.</p>
+
                         that the sgRNA might degraded.</div>
 
                     <img src="./img/group3May4pic1.png">
 
                     <img src="./img/group3May4pic1.png">
 
                     <img src="./img/group3May4pic2.png">
 
                     <img src="./img/group3May4pic2.png">
−
                     <p>Line1，plasmid；2，cleavage result</p>
+
                     <div class="notebook_week_p">Line1，plasmid；2，cleavage result</div>
 
                 </div>
 
                 </div>
  
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             <div class="notebook_Month_head">June</div>
 
             <div class="notebook_Month_head">June</div>
 
             <div class="notebook_week">
 
             <div class="notebook_week">
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                 <p style="text-align: left;font-size: 20px;text-indent: 2em;">Preparation for the final-term exams. </p>
+
                 <div style="text-align: left;font-size: 20px;text-indent: 2em;">Preparation for the final-term exams. </div>
  
 
             </div>
 
             </div>
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             <div class="notebook_Month_head">July</div>
 
             <div class="notebook_Month_head">July</div>
 
             <div class="notebook_week">
 
             <div class="notebook_week">
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                 <p style="text-align: left;font-size: 20px;text-indent: 0;">week1-3
+
                 <div style="text-align: left;font-size: 20px;text-indent: 0;">week1-3
 
                     <br> Most members joined a summer camp as a part of school lessons.
 
                     <br> Most members joined a summer camp as a part of school lessons.
 
                     <br> week4
 
                     <br> week4
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                     was used to remove RNase. But it turned out that quantity of DNA was not sufficient enough to be extracted.
 
                     was used to remove RNase. But it turned out that quantity of DNA was not sufficient enough to be extracted.
 
                     We tried for several times but the results of the nucleic acid gel electrophoresis showed that we got
 
                     We tried for several times but the results of the nucleic acid gel electrophoresis showed that we got
−
                     nothing after the purification.</p>
+
                     nothing after the purification.</div>
  
 
             </div>
 
             </div>
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                 </div>
 
                 </div>
 
                 <div id="8notebook_week1" class="notebook_information_word1" style="display:none;">
 
                 <div id="8notebook_week1" class="notebook_information_word1" style="display:none;">
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                     <p></p>
+
                      
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="8notebook_week2" class="notebook_information_word2" style="display:none;">
 
                 <div id="8notebook_week2" class="notebook_information_word2" style="display:none;">
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                     <p></p>
+
                      
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="8notebook_week3" class="notebook_information_word3" style="display:none;">
 
                 <div id="8notebook_week3" class="notebook_information_word3" style="display:none;">
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                     <p></p>
+
                      
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="8notebook_week4" class="notebook_information_word4" style="display:none;">
 
                 <div id="8notebook_week4" class="notebook_information_word4" style="display:none;">
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                     <p></p>
+
                      
 
                 </div>
 
                 </div>
  
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                 </div>
 
                 </div>
 
                 <div id="9notebook_week1" class="notebook_information_word1" style="display:none;">
 
                 <div id="9notebook_week1" class="notebook_information_word1" style="display:none;">
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                     <p></p>
+
                      
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="9notebook_week2" class="notebook_information_word2" style="display:none;">
 
                 <div id="9notebook_week2" class="notebook_information_word2" style="display:none;">
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                    <p></p>
+
                 
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="9notebook_week3" class="notebook_information_word3" style="display:none;">
 
                 <div id="9notebook_week3" class="notebook_information_word3" style="display:none;">
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                     <p></p>
+
                      
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="9notebook_week4" class="notebook_information_word4" style="display:none;">
 
                 <div id="9notebook_week4" class="notebook_information_word4" style="display:none;">
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                     <p></p>
+
                      
 
                 </div>
 
                 </div>
  
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                 </div>
 
                 </div>
 
                 <div id="10notebook_week1" class="notebook_information_word1" style="display:none;">
 
                 <div id="10notebook_week1" class="notebook_information_word1" style="display:none;">
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                     <p></p>
+
                      
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
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                 </div>
 
                 </div>
 
                 <div id="10notebook_week2" class="notebook_information_word2" style="display:none;">
 
                 <div id="10notebook_week2" class="notebook_information_word2" style="display:none;">
−
                     <p></p>
+
                      
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
Line 598: Line 598:
 
                 </div>
 
                 </div>
 
                 <div id="10notebook_week3" class="notebook_information_word3" style="display:none;">
 
                 <div id="10notebook_week3" class="notebook_information_word3" style="display:none;">
−
                     <p></p>
+
                      
 
                 </div>
 
                 </div>
 
                 <div>
 
                 <div>
Line 604: Line 604:
 
                 </div>
 
                 </div>
 
                 <div id="10notebook_week4" class="notebook_information_word4" style="display:none;">
 
                 <div id="10notebook_week4" class="notebook_information_word4" style="display:none;">
−
                     <p>test10.4</p>
+
                      
 
                 </div>
 
                 </div>
  
Line 644: Line 644:
 
                 })(i);
 
                 })(i);
 
             }
 
             }
−
 
+
            //点击隐藏显示
 
             function notebook_display(week, notebookweek) {
 
             function notebook_display(week, notebookweek) {
 
                 var p = document.getElementById(week);
 
                 var p = document.getElementById(week);

Revision as of 10:11, 17 October 2018

<!DOCTYPE >

class="notebook_month_p"
March
April
May
June
July
August
September
October
class="notebook_week_p"
March
April
May
June
Preparation for the final-term exams.
July
week1-3
Most members joined a summer camp as a part of school lessons.
week4
In order to reduce the influence of RNase on the subsequent experiment, the phenol-chloroform method was used to remove RNase. But it turned out that quantity of DNA was not sufficient enough to be extracted. We tried for several times but the results of the nucleic acid gel electrophoresis showed that we got nothing after the purification.
August
September
October