Difference between revisions of "Team:NJAU China/Notebook"

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<title>Team:NJAU_CHINA/Notebook </title>
  
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<h2>Notebook</h2>
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<p> Document the dates you worked on your project.</p>
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#MonthsTable{
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<h5>What should this page have?</h5>
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<ul>
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margin:3em 0;
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<li>Chronological notes of what your team is doing.</li>
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<li> Brief descriptions of daily important events.</li>
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#MonthsTable th,td{
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<li>Pictures of your progress. </li>
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<li>Mention who participated in what task.</li>
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th{
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<h4>Inspiration</h4>
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<p>You can see what others teams have done to organize their notes:</p>
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<ul>
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tr{
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<li><a href="https://2014.igem.org/Team:ATOMS-Turkiye/Notebook">2014 ATOMS-Turkiye</a></li>
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<li><a href="https://2014.igem.org/Team:Tec-Monterrey/ITESM14_project.html#tab_notebook">2014 Tec Monterrey</a></li>
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<li><a href="https://2014.igem.org/Team:Kyoto/Notebook/Magnetosome_Formation#title">2014 Kyoto</a></li>
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<li><a href="https://2014.igem.org/Team:Cornell/notebook">2014 Cornell</a></li>
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<div class="my_content">
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<h4>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Here is our experimental process.Filled out by each of our experiment teammates. The progress contains the details of our experiment, including the results of each step and the failure. And progress in our experiment can be clearly understood through this program.</h4>
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<table width="500" id="MonthsTable" cellspacing="0" cellpadding="0">
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<tr>
 +
<th>DATE</th>
 +
<th>LAB NOTE</th>
 +
<th>RESULT</th>
 +
</tr>
 +
<tr>
 +
<td>2015/3-4</td>
 +
<td>Discussed project</td>
 +
<td>Wonderful</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/5</td>
 +
<td>Made DH5α competent cell </td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/8</td>
 +
<td>TransformedBBa_R0082(Promoter (OmpR, positive))</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/10</td>
 +
<td>TransformedBBa_K592018 (cph8 with strong RB S),BBa_P0451(strongRBS:B0034+ cI (lambda): C0051+double terminator:B0010、B0012),BBa_C0040 (tetracycline repressor from transposon Tn10 (+LVA))</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/15</td>
 +
<td>TransformedBBa_K592016 (B0034-YF1-B0034-FixJ, blue light sensor and RR with RBS) , BBa_K823006(constitutive promoter) and BBa_K592006(FixK2 promoter)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/2-4/17</td>
 +
<td>Askedfor the cas9 part from other universities</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/10-18</td>
 +
<td>Askedfor the cas9 gene at our college and finally get from postgraduate</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/20</td>
 +
<td>Made competent cell DH5α</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/25</td>
 +
<td>TransformedBBa_S03518 (B0034-TetR);</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/28</td>
 +
<td>TransformedBBa_S03877(Strong RBS+hol), BBa_K081021(B003+Pcya+ terminator)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/4/29</td>
 +
<td>TransformedBBa_B0015 (Double terminator，forward： B0010+B0012)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/2</td>
 +
<td>Got Hfr straIn; PCR cas9 sequence and attempted to ligated cas9 on pSB1C3</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/3</td>
 +
<td>Made DH5αcompetent cell </td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/4</td>
 +
<td>PCR tested the existence of Orit and TraI gene in Hfr; Double restriction digestedBBa_S03877and BBa_K081021；ligateddigestedBBa_S03877 and digestedBBa_K081021</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/5</td>
 +
<td>Transformed the product of ligation on the former day into DH5α( the successful product was marked as holpt)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/7-8</td>
 +
<td>T-A colony of OriT from Hfr</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/10</td>
 +
<td>Double restriction digestedholpt and BBa_R0082;ligateddigestedholpt and BBa_R0082</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/11</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as holpto)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/11-12</td>
 +
<td>T-A colony of OriT from Hfr</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/14</td>
 +
<td>Test the conjugation efficiency between Hfr and BL21</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/19</td>
 +
<td>Tired to ligated cas9 with constitutive promoter ;Double restriction digestedBBa_S03518 and BBa_B0015 ;ligateddigestedBBa_S03518 and BBa_B0015</td>
 +
<td>Ligation failed; digestedion succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/20</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as tetT)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/24</td>
 +
<td>T-A colony of TraI from Hfr ; Double restriction digestedBBa_R0082 and holpto;ligateddigestedBBa_R0082 and holpto </td>
 +
<td>T-A colonyfailed; restriction digestedionsucceed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/25</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as cph8A)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/28</td>
 +
<td>Double restriction digestedtetT and BBa_C0040;ligateddigestedtetT and BBa_C0040</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/29</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as tetra)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/5/9-6/1</td>
 +
<td>Overlap PCR to knock out TraI gene in Hfr</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/2</td>
 +
<td>T-A colony of TraI from Hfr; TransformedBBa_I13504 (reporter  1.0RBS+GFP+Terminator)</td>
 +
<td>T-A colony failed; Transform edation succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/3</td>
 +
<td>Made DH5α competent cell </td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/4</td>
 +
<td>Double restriction digestedcph8A , tetra and BBa_I13504;ligated digeste dcph8A and BBa_I13504, , tetra and BBa_I13504</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/5</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as cpg and tetRg)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/9</td>
 +
<td>Double restriction digestedcph8A and tetRg;ligateddigestedcph8A and tetRg</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/10</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as ctg)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/12</td>
 +
<td>T-A colony of TraI from Hfr </td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/19</td>
 +
<td>Double restriction digestedBBa_K592016 (B0034-YF1-B0034-FixJ, blue light sensor and RR with RBS) and BBa_B0015 (Double terminator，forward： B0010+B0012); ligateddigestedBBa_K592016 and BBa_B0015</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/20</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as NT)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/20</td>
 +
<td>Made DH5αcompetent cell </td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/15-6/21</td>
 +
<td>DNA synthesis of promoter λ(the DNA is called Pci)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/25</td>
 +
<td>Double restriction digested NT,Pci , BBa_I13504and BBa_K592006;ligateddigested NT and BBa_K592006, Pci and BBa_K592006, Pci and BBa_I13504</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/26</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as NG,GCi and Cig)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/1</td>
 +
<td>Double restriction digested NG and BBa_K823006;ligateddigested NG and BBa_K823006</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/2</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as cngg)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/4</td>
 +
<td>Transformed the plasmid pET28a into DH5α</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/6</td>
 +
<td>Double restriction digestedpET28a and cngg；ligateddigestedpET28a and cngg</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/7</td>
 +
<td>Transformed the product of ligation on the former day into DH5α (the successful product was marked as pETcngg)</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/8</td>
 +
<td>Made BL21 competent cell </td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/10</td>
 +
<td>Transformed the plasmid pETcngg into Made BL21</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/13</td>
 +
<td>Double restriction digestedcpg and Pet28a;ligateddigestedcpg and Pet28a</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/14</td>
 +
<td>Transformed the product of ligation on the former day into DH5α</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/15</td>
 +
<td>Made DH5αcompetent cell </td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/6/5-7/15</td>
 +
<td>TraIgene knock out in Genescript company</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/17</td>
 +
<td>Transformed the product of ligation on the former day into DH5α</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/20</td>
 +
<td>Double restriction digestedcpg, ctg and BBa_K823006</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/16-7/20</td>
 +
<td>Test the conjugation efficiency between Hfr and BL21 and between TraIknocked-out Hfr and DH5α</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/21</td>
 +
<td>Transformed the product of ligation on the former day into DH5α</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/23</td>
 +
<td>Double restriction digestedcpg, ctg and BBa_K823006</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/24</td>
 +
<td>Transformed the product of ligation on the former day into DH5α</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/7/25-8/4</td>
 +
<td>Synthetized “GFPsg+terminater+constitutive promoter+1.0RBS” sequence by company.</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/5</td>
 +
<td>Made DH5αcompetent cell ; Ligatedconstitutive promoter with GFP</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/7</td>
 +
<td>Twice single restriction digestedcpg, ctg and Pet28a;ligateddigestedcpg and Pet28a; ctg and Pet28a; Ligated cas9 on pMD-18T</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/1-8/7</td>
 +
<td>Verified constructions of three plasmids needed for InterLab Study</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/8</td>
 +
<td>Transformed the product of ligation on the former day into DH5α</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/10</td>
 +
<td>Triedto ligate “GFPsg+terminater+ constitutive promoter+1.0RBS” sequence with cas9</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/10-8/12</td>
 +
<td>InterLab Study read florescence intensity: First attempt</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/13-8/15</td>
 +
<td>InterLab Study read florescence intensity: second attempt</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/15</td>
 +
<td>Double restriction digested cpg and Pet28a;ligated digested cpg and Pet28a</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/16</td>
 +
<td>Transformed the product of ligation on the former day into DH5α</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/20</td>
 +
<td>Ligatedcas9on pet-28a.</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/21</td>
 +
<td>Twice single restriction digested cpg, ctg and Pet28a;ligated digested cpg and Pet28a; ctg and Pet28a; Ligated cas9 on pMD-18T</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/22</td>
 +
<td>Transformed the product of ligation on the former day into DH5α</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/21</td>
 +
<td>T-A colony of TraI from Hfr</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/22</td>
 +
<td>Verification of T-A colony</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/23</td>
 +
<td>DNA sequencing of traI</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/24-27</td>
 +
<td>T-A colony of TraI from Hfr again</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/8/28</td>
 +
<td>Twice single restriction digested traI and pSB1C3; ligated traI and pSB1C3.</td>
 +
<td>Failed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/9/1-3</td>
 +
<td>Twice single restriction digested traI and pSB1C3; ligated traI and pSB1C3.</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/9/4-7</td>
 +
<td>Functional verification of traI</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/9/9</td>
 +
<td>Another T-A colony of OriT from Hfr</td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/9/10</td>
 +
<td>Made DH5αcompetent cell </td>
 +
<td>Succeed</td>
 +
</tr>
 +
<tr>
 +
<td>2015/9/10</td>
 +
<td>Submit the biobrick</td>
 +
<td>Succeed</td>
 +
</tr>
 +
</table>
 +
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Latest revision as of 14:07, 18 September 2015

Team:NJAU_CHINA/Notebook

     Here is our experimental process.Filled out by each of our experiment teammates. The progress contains the details of our experiment, including the results of each step and the failure. And progress in our experiment can be clearly understood through this program.

DATE LAB NOTE RESULT
2015/3-4 Discussed project Wonderful
2015/4/5 Made DH5α competent cell Succeed
2015/4/8 TransformedBBa_R0082(Promoter (OmpR, positive)) Succeed
2015/4/10 TransformedBBa_K592018 (cph8 with strong RB S),BBa_P0451(strongRBS:B0034+ cI (lambda): C0051+double terminator:B0010、B0012),BBa_C0040 (tetracycline repressor from transposon Tn10 (+LVA)) Succeed
2015/4/15 TransformedBBa_K592016 (B0034-YF1-B0034-FixJ, blue light sensor and RR with RBS) , BBa_K823006(constitutive promoter) and BBa_K592006(FixK2 promoter) Succeed
2015/4/2-4/17 Askedfor the cas9 part from other universities Failed
2015/4/10-18 Askedfor the cas9 gene at our college and finally get from postgraduate Succeed
2015/4/20 Made competent cell DH5α Succeed
2015/4/25 TransformedBBa_S03518 (B0034-TetR); Succeed
2015/4/28 TransformedBBa_S03877(Strong RBS+hol), BBa_K081021(B003+Pcya+ terminator) Succeed
2015/4/29 TransformedBBa_B0015 (Double terminator，forward： B0010+B0012) Succeed
2015/5/2 Got Hfr straIn; PCR cas9 sequence and attempted to ligated cas9 on pSB1C3 Succeed
2015/5/3 Made DH5αcompetent cell Succeed
2015/5/4 PCR tested the existence of Orit and TraI gene in Hfr; Double restriction digestedBBa_S03877and BBa_K081021；ligateddigestedBBa_S03877 and digestedBBa_K081021 Succeed
2015/5/5 Transformed the product of ligation on the former day into DH5α( the successful product was marked as holpt) Succeed
2015/5/7-8 T-A colony of OriT from Hfr Failed
2015/5/10 Double restriction digestedholpt and BBa_R0082;ligateddigestedholpt and BBa_R0082 Succeed
2015/5/11 Transformed the product of ligation on the former day into DH5α (the successful product was marked as holpto) Succeed
2015/5/11-12 T-A colony of OriT from Hfr Succeed
2015/5/14 Test the conjugation efficiency between Hfr and BL21 Succeed
2015/5/19 Tired to ligated cas9 with constitutive promoter ;Double restriction digestedBBa_S03518 and BBa_B0015 ;ligateddigestedBBa_S03518 and BBa_B0015 Ligation failed; digestedion succeed
2015/5/20 Transformed the product of ligation on the former day into DH5α (the successful product was marked as tetT) Succeed
2015/5/24 T-A colony of TraI from Hfr ; Double restriction digestedBBa_R0082 and holpto;ligateddigestedBBa_R0082 and holpto T-A colonyfailed; restriction digestedionsucceed
2015/5/25 Transformed the product of ligation on the former day into DH5α (the successful product was marked as cph8A) Succeed
2015/5/28 Double restriction digestedtetT and BBa_C0040;ligateddigestedtetT and BBa_C0040 Succeed
2015/5/29 Transformed the product of ligation on the former day into DH5α (the successful product was marked as tetra) Succeed
2015/5/9-6/1 Overlap PCR to knock out TraI gene in Hfr Failed
2015/6/2 T-A colony of TraI from Hfr; TransformedBBa_I13504 (reporter 1.0RBS+GFP+Terminator) T-A colony failed; Transform edation succeed
2015/6/3 Made DH5α competent cell Succeed
2015/6/4 Double restriction digestedcph8A , tetra and BBa_I13504;ligated digeste dcph8A and BBa_I13504, , tetra and BBa_I13504 Succeed
2015/6/5 Transformed the product of ligation on the former day into DH5α (the successful product was marked as cpg and tetRg) Succeed
2015/6/9 Double restriction digestedcph8A and tetRg;ligateddigestedcph8A and tetRg Succeed
2015/6/10 Transformed the product of ligation on the former day into DH5α (the successful product was marked as ctg) Succeed
2015/6/12 T-A colony of TraI from Hfr Failed
2015/6/19 Double restriction digestedBBa_K592016 (B0034-YF1-B0034-FixJ, blue light sensor and RR with RBS) and BBa_B0015 (Double terminator，forward： B0010+B0012); ligateddigestedBBa_K592016 and BBa_B0015 Succeed
2015/6/20 Transformed the product of ligation on the former day into DH5α (the successful product was marked as NT) Succeed
2015/6/20 Made DH5αcompetent cell Succeed
2015/6/15-6/21 DNA synthesis of promoter λ(the DNA is called Pci) Succeed
2015/6/25 Double restriction digested NT,Pci , BBa_I13504and BBa_K592006;ligateddigested NT and BBa_K592006, Pci and BBa_K592006, Pci and BBa_I13504 Succeed
2015/6/26 Transformed the product of ligation on the former day into DH5α (the successful product was marked as NG,GCi and Cig) Succeed
2015/7/1 Double restriction digested NG and BBa_K823006;ligateddigested NG and BBa_K823006 Succeed
2015/7/2 Transformed the product of ligation on the former day into DH5α (the successful product was marked as cngg) Succeed
2015/7/4 Transformed the plasmid pET28a into DH5α Succeed
2015/7/6 Double restriction digestedpET28a and cngg；ligateddigestedpET28a and cngg Succeed
2015/7/7 Transformed the product of ligation on the former day into DH5α (the successful product was marked as pETcngg) Succeed
2015/7/8 Made BL21 competent cell Succeed
2015/7/10 Transformed the plasmid pETcngg into Made BL21 Succeed
2015/7/13 Double restriction digestedcpg and Pet28a;ligateddigestedcpg and Pet28a Succeed
2015/7/14 Transformed the product of ligation on the former day into DH5α Failed
2015/7/15 Made DH5αcompetent cell Succeed
2015/6/5-7/15 TraIgene knock out in Genescript company Succeed
2015/7/17 Transformed the product of ligation on the former day into DH5α Failed
2015/7/20 Double restriction digestedcpg, ctg and BBa_K823006 Succeed
2015/7/16-7/20 Test the conjugation efficiency between Hfr and BL21 and between TraIknocked-out Hfr and DH5α Succeed
2015/7/21 Transformed the product of ligation on the former day into DH5α Failed
2015/7/23 Double restriction digestedcpg, ctg and BBa_K823006 Succeed
2015/7/24 Transformed the product of ligation on the former day into DH5α Failed
2015/7/25-8/4 Synthetized “GFPsg+terminater+constitutive promoter+1.0RBS” sequence by company. Succeed
2015/8/5 Made DH5αcompetent cell ; Ligatedconstitutive promoter with GFP Succeed
2015/8/7 Twice single restriction digestedcpg, ctg and Pet28a;ligateddigestedcpg and Pet28a; ctg and Pet28a; Ligated cas9 on pMD-18T Succeed
2015/8/1-8/7 Verified constructions of three plasmids needed for InterLab Study Succeed
2015/8/8 Transformed the product of ligation on the former day into DH5α Failed
2015/8/10 Triedto ligate “GFPsg+terminater+ constitutive promoter+1.0RBS” sequence with cas9 Failed
2015/8/10-8/12 InterLab Study read florescence intensity: First attempt Failed
2015/8/13-8/15 InterLab Study read florescence intensity: second attempt Succeed
2015/8/15 Double restriction digested cpg and Pet28a;ligated digested cpg and Pet28a Succeed
2015/8/16 Transformed the product of ligation on the former day into DH5α Failed
2015/8/20 Ligatedcas9on pet-28a. Failed
2015/8/21 Twice single restriction digested cpg, ctg and Pet28a;ligated digested cpg and Pet28a; ctg and Pet28a; Ligated cas9 on pMD-18T Succeed
2015/8/22 Transformed the product of ligation on the former day into DH5α Failed
2015/8/21 T-A colony of TraI from Hfr Succeed
2015/8/22 Verification of T-A colony Succeed
2015/8/23 DNA sequencing of traI Failed
2015/8/24-27 T-A colony of TraI from Hfr again Succeed
2015/8/28 Twice single restriction digested traI and pSB1C3; ligated traI and pSB1C3. Failed
2015/9/1-3 Twice single restriction digested traI and pSB1C3; ligated traI and pSB1C3. Succeed
2015/9/4-7 Functional verification of traI Succeed
2015/9/9 Another T-A colony of OriT from Hfr Succeed
2015/9/10 Made DH5αcompetent cell Succeed
2015/9/10 Submit the biobrick Succeed