Team:HokkaidoU Japan/Notebook/asB0034/asB0034-construction
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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 02:46, 18 October 2014
Notebook
Lab Documents
asB0034 Construction
-
Start
-
Primer Annealing
asB0034_F Tm:68.7℃ asB0034_R Tm:68.7℃ -
Digestion
Cut pHN1257 with NcoI, XhoI(using 10×cut smart) -
Gel extraction
digested pHN1257 -
Ethanol Precipetation
digested pHN1257 -
Ligation
Ligate pHN1257 with asB0034 -
Transformation
Ptrc-stem-asB0034-stem on pHN1257 5.0µL DNA to DH5αColony PCR
Ptrc-stem-asB0034-stem(on pHN1257) pHN1257_up_F Tm:60.1℃ & asB0034-R Tm:68.7℃Culture
Ptrc-stem-asB0034-stem(on pHN1257)mini-Prep
Ptrc-stem-asB0034-stem(on pHN1257)Double Transformation
Ptrc-stem-asB0034-stem(on pHN1257)&R0040-B0034-E1010-B0015(on pSB6A1) each of 2.5µL DNA to DH5αComplete!