Team:HokkaidoU Japan/Notebook/asB0034/asB0034,32-1C3-construction

From 2014.igem.org

(Difference between revisions)
Line 140: Line 140:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
Cut  vector with EcoRI, PstI (using 10&times;H buffer)  Cut  insert with EcoRI, PstI (using 10&times;M buffer)
+
Cut  vector with EcoRI, PstI (using 10&times;H buffer)   
 +
</div>
 +
<div class="step-recipe">
 +
Cut  insert with EcoRI, PstI (using 10&times;M buffer)
</div>
</div>
    </div>
    </div>
Line 245: Line 248:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
ligation products 5.0&micro;L to DH5&alpha;
+
ligation products
 +
</div>
 +
<div class="step-recipe">
 +
5.0&micro;L to DH5&alpha;
</div>
</div>
    </div>
    </div>
Line 266: Line 272:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
R0010-stem-asB0034-stem-B0015(on pSB1C3) 100up_F Tm:62.6&#8451; & asB0034_R Tm:68.7&#8451; R0010-stem-asB0032-stem-B0015(on pSB1C3) 100up_F Tm:62.6&#8451; & asB0032_R Tm:70.8&#8451;
+
R0010-stem-asB0034-stem-B0015(on pSB1C3) 100up_F Tm:62.6&#8451; & asB0034_R Tm:68.7&#8451;  
 +
</div>
 +
<div class="step-recipe">
 +
R0010-stem-asB0032-stem-B0015(on pSB1C3) 100up_F Tm:62.6&#8451; & asB0032_R Tm:70.8&#8451;
</div>
</div>
    </div>
    </div>
Line 287: Line 296:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
R0010-stem-asB0034-stem-B0015(on pSB1C3) R0010-stem-asB0032-stem-B0015(on pSB1C3)
+
R0010-stem-asB0034-stem-B0015(on pSB1C3)  
 +
</div>
 +
<div class="step-recipe">
 +
R0010-stem-asB0032-stem-B0015(on pSB1C3)
</div>
</div>
    </div>
    </div>
Line 308: Line 320:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
R0010-stem-asB0034-stem-B0015(on pSB1C3) R0010-stem-asB0032-stem-B0015(on pSB1C3)
+
R0010-stem-asB0034-stem-B0015(on pSB1C3)  
 +
</div>
 +
<div class="step-recipe">
 +
R0010-stem-asB0032-stem-B0015(on pSB1C3)
</div>
</div>
    </div>
    </div>
Line 329: Line 344:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
R0010-stem-asB0034-stem-B0015(on pSB1C3) with 100up_F Tm:62.8&#8451; & 200dn:66.6&#8451; R0010-stem-asB0032-stem-B0015(on pSB1C3) with 100up_F Tm:62.6&#8451; 200dn_R Tm:66.6&#8451;
+
R0010-stem-asB0034-stem-B0015(on pSB1C3) with 100up_F Tm:62.8&#8451; & 200dn_R Tm:66.6&#8451;
 +
</div>
 +
<div class="step-recipe">
 +
R0010-stem-asB0032-stem-B0015(on pSB1C3) with 100up_F Tm:62.6&#8451; & 200dn_R Tm:66.6&#8451;
</div>
</div>
    </div>
    </div>

Revision as of 03:10, 18 October 2014

Notebook
Lab Documents

Anti-Sense B0034,B0032 on 1C3 Construction

  • Start

  • Digestion

    Cut vector with EcoRI, PstI (using 10×H buffer)
    Cut insert with EcoRI, PstI (using 10×M buffer)

  • Digestion

    Cut vector with EcoRI, PstI (using 10×H buffer)

  • Gel extraction

    digested vector insert

  • Ethanol Precipitation

    digested vector insert

  • Ligation

    Ligate vector with insert

  • Transformation

    ligation products
    5.0µL to DH5α

  • Colony PCR

    R0010-stem-asB0034-stem-B0015(on pSB1C3) 100up_F Tm:62.6℃ & asB0034_R Tm:68.7℃
    R0010-stem-asB0032-stem-B0015(on pSB1C3) 100up_F Tm:62.6℃ & asB0032_R Tm:70.8℃

  • Culture

    R0010-stem-asB0034-stem-B0015(on pSB1C3)
    R0010-stem-asB0032-stem-B0015(on pSB1C3)

  • Mini-Prep

    R0010-stem-asB0034-stem-B0015(on pSB1C3)
    R0010-stem-asB0032-stem-B0015(on pSB1C3)

  • Sequencing

    R0010-stem-asB0034-stem-B0015(on pSB1C3) with 100up_F Tm:62.8℃ & 200dn_R Tm:66.6℃
    R0010-stem-asB0032-stem-B0015(on pSB1C3) with 100up_F Tm:62.6℃ & 200dn_R Tm:66.6℃

  • Complete!

Retrieved from "http://2014.igem.org/Team:HokkaidoU_Japan/Notebook/asB0034/asB0034,32-1C3-construction"