Team:SJTU-BioX-Shanghai/daynotes

From 2014.igem.org

(Difference between revisions)
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                 <article class="post__article">
                     <h2 id ="JulyWeek1" >July Week1</h2>
                     <h2 id ="JulyWeek1" >July Week1</h2>
-
                     <p>Plasmid Amplification for further construction; chose pRSFDuet-1, pACYCDuet-1 and pCDFDuet-1 for expression vector and pBluescript II KS(+) for connector.<p>
+
                     <p>Plasmid Amplification for further construction;  
 +
                        chose pRSFDuet-1, pACYCDuet-1 and pCDFDuet-1 for expression vector and pBluescript II KS(+) for connector.<p>
                     <h2 id ="JulyWeek2" >July Week2</h2>
                     <h2 id ="JulyWeek2" >July Week2</h2>
-
                     <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>
+
                     <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>
                     <h2 id ="JulyWeek3" >July Week3</h2>
                     <h2 id ="JulyWeek3" >July Week3</h2>
-
                     <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>
+
                     <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>
                     <h2 id ="JulyWeek4" >July Week4</h2>
                     <h2 id ="JulyWeek4" >July Week4</h2>
-
                     <p> Use golden gate to connected TAL of 2012 Freiburg igem. Transform, colony picking plasmid extraction and digestion identification.<p>  
+
                     <p> Use golden gate to connected TAL of 2012 Freiburg igem.  
 +
                          Transform, colony picking plasmid extraction and digestion identification.<p>  
                     <h2 id ="AugustWeek1-2" >August Week1-2</h2>
                     <h2 id ="AugustWeek1-2" >August Week1-2</h2>
-
                     <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>
+
                     <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>
                     <h2 id ="AugustWeek3" >August Week3</h2>
-
                     <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>
+
                     <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>
                     <h2 id ="AugustWeek4" >August Week4</h2>
-
                     <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>     
+
                     <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 ="SeptemberWeek1" >September Week1</h2>
                     <h2 id ="SeptemberWeek1" >September Week1</h2>
-
                     <p>Try co-transformation: Prsf  pacyc  pBluescript . Find the conditions of protein expression. Find the way to construct the TAL.<p>           
+
                     <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>
                     <h2 id ="SeptemberWeek2" >September Week2</h2>
-
                     <p>Find the enzymes for the application. Find the way to detect the substrate in these pathways. Connector plasmid modification.<p>     
+
                     <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>
                     <h2 id ="SeptemberWeek3" >September Week3</h2>
                     <p>TAL gene synthesis. Construct the part with our new ports.<p>  
                     <p>TAL gene synthesis. Construct the part with our new ports.<p>  

Revision as of 14:22, 16 October 2014

July Week1

Plasmid Amplification for further construction; chose pRSFDuet-1, pACYCDuet-1 and pCDFDuet-1 for expression vector and pBluescript II KS(+) for connector.

July Week2

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.

July Week3

Connected ssDsbA-FP-HL-lgt by splicing by overlap extension. Transform, colony picking plasmid extraction and digestion identification; Sequencing results showed accurate construction.

July Week4

Use golden gate to connected TAL of 2012 Freiburg igem. Transform, colony picking plasmid extraction and digestion identification.

August Week1-2

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.

August Week3

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.

August Week4

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.

September Week1

Try co-transformation: Prsf pacyc pBluescript . Find the conditions of protein expression. Find the way to construct the TAL.

September Week2

Find the enzymes for the application. Find the way to detect the substrate in these pathways. Connector plasmid modification.

September Week3

TAL gene synthesis. Construct the part with our new ports.

September Week4

TAL gene synthesis.