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Team:HokkaidoU Japan/Notebook/asB0034/asB0034,32-construction
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{{Team:HokkaidoU_Japan/Footer/CSS}} | {{Team:HokkaidoU_Japan/Footer/CSS}} | ||
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<div class="box"> | <div class="box"> | ||
<div id="box-wrapper"> | <div id="box-wrapper"> | ||
| - | |||
<ul id="ldd_menu" class="ldd_menu"> | <ul id="ldd_menu" class="ldd_menu"> | ||
<li> | <li> | ||
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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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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | asB0034_F Tm: 68.7℃ & asB0034_R Tm: 68.7℃ asB0032_F Tm: 71.6℃ & asB0032_R Tm: 70.8℃ | + | asB0034_F Tm: 68.7℃ & asB0034_R Tm: 68.7℃ |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | asB0032_F Tm: 71.6℃ & asB0032_R Tm: 70.8℃ | ||
</div> | </div> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ligate pSB1A3 | + | Ligate pSB1A3 with R0010-stem-asB0034-stem-B0015 |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | Ligate pSB1A3 with R0010-stem-asB0032-stem-B0015 | ||
</div> | </div> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | ligation products 5.0µL to DH5α | + | ligation products |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | 5.0µL to DH5α | ||
</div> | </div> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem-asB0034-stem-B0015(on pSB1A3) 100up_F Tm:62.8℃ & asB0034_R Tm:68.7℃ R0010-stem-asB0032-stem-B0015(on pSB1A3) 100up_F Tm:74.5℃ & asB0032_R Tm:70.8℃ Kapa-Taq | + | R0010-stem-asB0034-stem-B0015(on pSB1A3) 100up_F Tm:62.8℃ & asB0034_R Tm:68.7℃ |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | R0010-stem-asB0032-stem-B0015(on pSB1A3) 100up_F Tm:74.5℃ & asB0032_R Tm:70.8℃ | ||
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | Kapa-Taq | ||
</div> | </div> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem-asB0034-stem-B0015(on pSB1A3) R0010-stem-asB0032-stem-B0015(on pSB1A3) | + | R0010-stem-asB0034-stem-B0015(on pSB1A3) |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | R0010-stem-asB0032-stem-B0015(on pSB1A3) | ||
</div> | </div> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem-asB0034-stem-B0015(on pSB1A3) R0010-stem-asB0032-stem-B0015(on pSB1A3) | + | R0010-stem-asB0034-stem-B0015(on pSB1A3) |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | R0010-stem-asB0032-stem-B0015(on pSB1A3) | ||
</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 15:29, 9 September 2015
Notebook
Lab Documents
Anti-Sense B0034 (B0032) Construction
-
Start
-
Primer Annealing
asB0034_F Tm: 68.7℃ & asB0034_R Tm: 68.7℃asB0032_F Tm: 71.6℃ & asB0032_R Tm: 70.8℃ -
Digestion
Cut vector with NcoI, XhoI (using 10×cut smart buffer) -
Gel extraction
digested pSB1A3 -
Ethanol Precipitation
digested pSB1A3 -
Ligation
Ligate pSB1A3 with R0010-stem-asB0034-stem-B0015Ligate pSB1A3 with R0010-stem-asB0032-stem-B0015 -
Transformation
ligation products5.0µL to DH5α -
Colony PCR
R0010-stem-asB0034-stem-B0015(on pSB1A3) 100up_F Tm:62.8℃ & asB0034_R Tm:68.7℃R0010-stem-asB0032-stem-B0015(on pSB1A3) 100up_F Tm:74.5℃ & asB0032_R Tm:70.8℃Kapa-Taq -
Liquid Culture
R0010-stem-asB0034-stem-B0015(on pSB1A3)R0010-stem-asB0032-stem-B0015(on pSB1A3) -
Mini-Prep
R0010-stem-asB0034-stem-B0015(on pSB1A3)R0010-stem-asB0032-stem-B0015(on pSB1A3) -
Complete!
