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Team:HokkaidoU Japan/Notebook/Pre experiment/asmRFP
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> | ||
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<div class="step-tile"> | <div class="step-tile"> | ||
<h2 class="step-title"> | <h2 class="step-title"> | ||
| - | Get anti-sense vector | + | Get anti-sense vector |
</h2> | </h2> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0040-B0034-E1010-B0015 with as_RBS_XhoI_F Tm:68.2℃ & as_mRFP_NcoI_R Tm:76.0℃ KOD plus Neo | + | R0040-B0034-E1010-B0015 with as_RBS_XhoI_F Tm:68.2℃ & as_mRFP_NcoI_R Tm:76.0℃ |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | KOD plus Neo | ||
</div> | </div> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Cut pHN1257 with NcoI, XhoI (using 10×Cut Smart) Cut asmRFP with NcoI, XhoI (using 10×Cut Smart) | + | Cut pHN1257 with NcoI, XhoI (using 10×Cut Smart) |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | Cut asmRFP with NcoI, XhoI (using 10×Cut Smart) | ||
</div> | </div> | ||
</div> | </div> | ||
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<div class="step-tile"> | <div class="step-tile"> | ||
<h2 class="step-title"> | <h2 class="step-title"> | ||
| - | Transformation | + | Transformation |
</h2> | </h2> | ||
</div> | </div> | ||
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<li> | <li> | ||
<time> | <time> | ||
| - | + | Sat Aug 16 | |
</time> | </time> | ||
</li> | </li> | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | asmRFP(on pHN1257) 5µL to DH5α Turbo | + | asmRFP(on pHN1257) |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | 5µL to DH5α Turbo | ||
</div> | </div> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | asmRFP(on pHN1257) pHN1257_up_F Tm:60.1℃ & pHN1257_terminator_R Tm:71.4℃ Kapa-Taq | + | asmRFP(on pHN1257) pHN1257_up_F Tm:60.1℃ & pHN1257_terminator_R Tm:71.4℃ |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | Kapa-Taq | ||
</div> | </div> | ||
</div> | </div> | ||
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<div id="footer-content"> | <div id="footer-content"> | ||
<div id="footer-logo"> | <div id="footer-logo"> | ||
| - | <a href="http://igemhokkaidou.com | + | <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 15:23, 9 September 2015
Notebook
Lab Documents
Anti-Sense mRFP Construction
-
Start
-
Get anti-sense vector
Great thanks to N. Nakashima -
PCR & PCR purification
R0040-B0034-E1010-B0015 with as_RBS_XhoI_F Tm:68.2℃ & as_mRFP_NcoI_R Tm:76.0℃KOD plus Neo -
Ethanol Precipitation
asmRFP -
Digestion
Cut pHN1257 with NcoI, XhoI (using 10×Cut Smart)Cut asmRFP with NcoI, XhoI (using 10×Cut Smart) -
Gel Extraction
pHN1257 asmRFP -
Ethanol Precipitation
asmRFP -
Dephosphorylation
asmRFP -
Ligation
Ligate asmRFP with pHN1257 -
Transformation
asmRFP(on pHN1257)5µL to DH5α Turbo -
Colony PCR
asmRFP(on pHN1257) pHN1257_up_F Tm:60.1℃ & pHN1257_terminator_R Tm:71.4℃Kapa-Taq -
Liquid Culture
asmRFP(on pHN1257) -
Mini-Prep
asmRFP(pHN1257) -
Complete!
