Line 62: | Line 62: | ||
</div> <br /><br /><br /><br /><br /><br /> | </div> <br /><br /><br /><br /><br /><br /> | ||
− | <section><h3>1. Cloning of module 1:</h3><h4>Construct 1--> Ptet - RBS1 - T7 RNA polymerase - terminator 1 - terminator 2. | + | <section><h3>1. Cloning of module 1:</h3><h4>Construct 1 --> Ptet - RBS1 - T7 RNA polymerase - terminator 1 - terminator 2. |
Two PCR reactions were performed. The first PCR reaction was performed with primer 1 and 2 using the template R0040 to amplify the Ptet. The size of PCR product is 77 bp. The second PCR reaction was performed with primer 3 and 4 using the template K1450004 to amplify the RBS1 - T7 RNA polymerase - terminator 1 - terminator 2. The size of this PCR product is 2338 bp. | Two PCR reactions were performed. The first PCR reaction was performed with primer 1 and 2 using the template R0040 to amplify the Ptet. The size of PCR product is 77 bp. The second PCR reaction was performed with primer 3 and 4 using the template K1450004 to amplify the RBS1 - T7 RNA polymerase - terminator 1 - terminator 2. The size of this PCR product is 2338 bp. | ||
Thirdly, SOE PCR was performed using the primers 1 and 4 (for PCR conditions and reaction mixture, refer notebook). | Thirdly, SOE PCR was performed using the primers 1 and 4 (for PCR conditions and reaction mixture, refer notebook). | ||
Line 81: | Line 81: | ||
<center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/2/26/T--IISER-Mohali-INDIA--results1.jpg" alt="Screening of transformants containing construct 2 by resriction digestion with SalI and BamHI"></td></tr></table></center> | <center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/2/26/T--IISER-Mohali-INDIA--results1.jpg" alt="Screening of transformants containing construct 2 by resriction digestion with SalI and BamHI"></td></tr></table></center> | ||
<h3>2. Cloning of module 2:</h3> | <h3>2. Cloning of module 2:</h3> | ||
− | <h4>Construct 1--> Pmar - RBS4 - ToxR - terminator 4.</h4> | + | <h4>Construct 1 --> Pmar - RBS4 - ToxR - terminator 4.</h4> |
<h4>The first PCR was performed with primers 11 and 12 using the template <i>E. coli</i> genome to amplify the pmar - RBS4. The size of the PCR product is 524 bp. | <h4>The first PCR was performed with primers 11 and 12 using the template <i>E. coli</i> genome to amplify the pmar - RBS4. The size of the PCR product is 524 bp. | ||
The second PCR was performed with primers 13 and 14 using the template K641009 to amplify ToxR. The size of this PCR product is 1441 bp. | The second PCR was performed with primers 13 and 14 using the template K641009 to amplify ToxR. The size of this PCR product is 1441 bp. | ||
Line 94: | Line 94: | ||
<center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/7/72/T--IISER-Mohali-INDIA--rslstss.png" alt="Flow chart"></td></tr></table></center> | <center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/7/72/T--IISER-Mohali-INDIA--rslstss.png" alt="Flow chart"></td></tr></table></center> | ||
<center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/3/31/T--IISER-Mohali-INDIA--CS4.png" alt="Flow chart"></td></tr></table></center> | <center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/3/31/T--IISER-Mohali-INDIA--CS4.png" alt="Flow chart"></td></tr></table></center> | ||
− | <h4>Construct 2--> Pctx - RBS5 - chromoprotein I - RBS6 - TetR - terminator5.</h4> | + | <h4>Construct 2 --> Pctx - RBS5 - chromoprotein I - RBS6 - TetR - terminator5.</h4> |
<center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/6/63/T--IISER-Mohali-INDIA--CS5.png" alt="Flow chart"></td></tr></table></center> | <center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/6/63/T--IISER-Mohali-INDIA--CS5.png" alt="Flow chart"></td></tr></table></center> | ||
<h4>Two PCR reactions were performed. The first PCR reaction was performed with primers 17 and 18 to amplify the Pctx. The size of PCR product is 180 bp. | <h4>Two PCR reactions were performed. The first PCR reaction was performed with primers 17 and 18 to amplify the Pctx. The size of PCR product is 180 bp. |
Revision as of 00:08, 2 November 2017