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| | </div> | | </div> |
| | <div class="content"> | | <div class="content"> |
| - | <p><a href="#a">Molecule</a></p> | + | <p><a href="https://2014.igem.org/Team:SJTU-BioX-Shanghai/Protocol#a">Molecule</a></p> |
| - | <p><a href="#b">Cell</a></p> | + | <p><a href="https://2014.igem.org/Team:SJTU-BioX-Shanghai/Protocol#b">Cell</a></p> |
| - | <p><a href="#d">Protein</a></p> | + | <p><a href="https://2014.igem.org/Team:SJTU-BioX-Shanghai/Protocol#d">Protein</a></p> |
| | </div> | | </div> |
| | </div> | | </div> |
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| | <div class="content"> | | <div class="content"> |
| | <article class="post__article"> | | <article class="post__article"> |
| - | <h2 id="July">July Week1</h2><br> | + | <h2 id="Molecule">Molecule</h2> |
| | <p>Plasmid Amplification for further construction; | | <p>Plasmid Amplification for further construction; |
| | <br>chose pRSFDuet-1, pACYCDuet-1 and pCDFDuet-1 for expression vector and pBluescript II KS（+） for connector.<p> | | <br>chose pRSFDuet-1, pACYCDuet-1 and pCDFDuet-1 for expression vector and pBluescript II KS（+） for connector.<p> |
| - | <h2 id ="JulyWeek2" >July Week2</h2><br> | + | <h2 id ="Cell" >Cell</h2> |
| | <p>Construct the gene of our fusion protein: ssDsbA-FP-HL-Lgt-FL-TAL using splicing by overlap extension. | | <p>Construct the gene of our fusion protein: ssDsbA-FP-HL-Lgt-FL-TAL using splicing by overlap extension. |
| | We connected ssDsbA-FP-HL-lgt for one part of this fusion protein and FL-TAL for another. Then connect them together.<p> | | We connected ssDsbA-FP-HL-lgt for one part of this fusion protein and FL-TAL for another. Then connect them together.<p> |
| - | <h2 id ="JulyWeek3" >July Week3</h2><br> | + | <h2 id ="Protein" >Protein</h2> |
| | <p> Connected ssDsbA-FP-HL-lgt by splicing by overlap extension. | | <p> Connected ssDsbA-FP-HL-lgt by splicing by overlap extension. |
| - | Transform, colony picking plasmid extraction and digestion identification; Sequencing results showed accurate construction.<p> | + | Transform, colony picking plasmid extraction and digestion identification; Sequencing results showed accurate construction.<p> |
| - | <h2 id ="JulyWeek4" >July Week4</h2><br>
| + | |
| - | <p> Use golden gate to connected TAL of 2012 Freiburg igem.
| + | |
| - | Transform, colony picking plasmid extraction and digestion identification.<p>
| + | |
| - | <h2 id="August">August Week1-2</h2><br>
| + | |
| - | <p>Test the conditions for the PCR.
| + | |
| - | Connected TAL, transform, colony picking plasmid extraction and digestion identification. Find with our electrophoresis band. Expression vectors and connector plasmid are confirmed by sequencing.<p>
| + | |
| - | <h2 id ="AugustWeek3" >August Week3</h2><br>
| + | |
| - | <p>There are some problems about Freiburg’s parts. We can’t connected TAL in the right order.
| + | |
| - | So we design some new primes for PCR that can produce the right sequence.<p>
| + | |
| - | <h2 id ="AugustWeek4" >August Week4</h2><br>
| + | |
| - | <p>Design a few new ports for the fusion protein.
| + | |
| - | Sequencing results showed accurate construction. Observe the FP using LSCM to confirm the fusion protein can locate on the membrane.<p>
| + | |
| - | <h2 id="September" >September Week1</h2><br>
| + | |
| - | <p>Try co-transformation: Prsf pacyc pBluescript .
| + | |
| - | Find the conditions of protein expression.
| + | |
| - | Find the way to construct the TAL.<p>
| + | |
| - | <h2 id ="SeptemberWeek2" >September Week2</h2><br>
| + | |
| - | <p>Find the enzymes for the application.
| + | |
| - | Find the way to detect the substrate in these pathways.
| + | |
| - | Connector plasmid modification.<p>
| + | |
| - | <h2 id ="SeptemberWeek3" >September Week3</h2><br>
| + | |
| - | <p>TAL gene synthesis. Construct the part with our new ports.<p>
| + | |
| - | <h2 id ="SeptemberWeek4" >September Week4</h2><br>
| + | |
| - | <p> TAL gene synthesis.<p>
| + | |
| | </article> | | </article> |
| | </div> | | </div> |
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