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Team:HokkaidoU Japan/Notebook/asB0034/asB0034,32-1C3-construction
From 2014.igem.org
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<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# | + | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Evaluation">Evaluation</a></li> |
</ul> | </ul> | ||
</div> | </div> | ||
Revision as of 02:50, 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:66.6℃ R0010-stem-asB0032-stem-B0015(on pSB1C3) with 100up_F Tm:62.6℃ 200dn_R Tm:66.6℃ -
Complete!
