Team:HokkaidoU Japan/Notebook/Pre experiment/mRFP-expression-construct

From 2014.igem.org

(Difference between revisions)
 
(2 intermediate revisions not shown)
Line 89: Line 89:
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion">Discussion</a></li>
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion">Discussion</a></li>
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Background">Background</a></li>
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Background">Background</a></li>
-
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Estimation">Evaluation</a></li>
+
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Evaluation">Evaluation</a></li>
</ul>
</ul>
</div>
</div>
Line 194: Line 194:
      <div class="step-recipe"> R0040-B0034-E1010-B0015 from our stock
      <div class="step-recipe"> R0040-B0034-E1010-B0015 from our stock
100up_F Tm:70.4&#8451; & 200dn_R Tm:66.6&#8451;
100up_F Tm:70.4&#8451; & 200dn_R Tm:66.6&#8451;
 +
</div>
 +
<div class="step-recipe">
         KOD plus Neo</div>
         KOD plus Neo</div>
    </div>
    </div>
Line 210: Line 212:
      </ul>
      </ul>
      <div class="step-recipe"> Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10&times;H buffer)
      <div class="step-recipe"> Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10&times;H buffer)
 +
</div>
 +
<div class="step-recipe">
Cut pSB6A1 with EcoRI, PstI (using 10&times;H buffer)
Cut pSB6A1 with EcoRI, PstI (using 10&times;H buffer)
</div>
</div>
Line 341: Line 345:
   <div id="footer-content">
   <div id="footer-content">
<div id="footer-logo">
<div id="footer-logo">
-
<a href="http://igemhokkaidou.com/"><img style="height:150px;position:relative;" src="https://static.igem.org/mediawiki/2014/3/39/HokkaidoU_logo_transparent.png"></a>
+
<a href="http://igemhokkaidou.wordpress.com"><img style="height:150px;position:relative;" src="https://static.igem.org/mediawiki/2014/3/39/HokkaidoU_logo_transparent.png"></a>
</div>
</div>
<div id="footer-twitter">
<div id="footer-twitter">

Latest revision as of 15:24, 9 September 2015

Notebook
Lab Documents

mRFP Expression Plasmid Construction

  • Start

  • Get genes

    pSB6A1
  • Transformation

    pSB6A1 1µL to JM109
  • Liquid Culture

    pSB6A1
  • Mini-prep

    pSB6A1
  • PCR & PCR purification

    R0040-B0034-E1010-B0015 from our stock 100up_F Tm:70.4℃ & 200dn_R Tm:66.6℃
    KOD plus Neo
  • Digestion

    Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10×H buffer)
    Cut pSB6A1 with EcoRI, PstI (using 10×H buffer)
  • Gel Extraction

    R0040-B0034-E1010-B0015 pSB6A1
  • Ethanol precipetation

    R0040-B0034-E1010-B0015 pSB6A1
  • Dephosphorylation

    R0040-B0034-E1010-B0015
  • Ligation

    Ligate R0040-B0034-E1010-B0015 with pSB6A1
  • Transformation

    R0040-B0034-E1010-B0015(on pSB6A1) 5µL to DH5α Turbo
  • Liquid Culture

    R0040-B0034-E1010-B0015(on pSB6A1)
  • Mini-prep

    R0040-B0034-E1010-B0015(on pSB6A1)
  • Complete!