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Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement
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<div id="header-bottom-contents"> | <div id="header-bottom-contents"> | ||
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| + | <div class="header-title-top"> | ||
| + | Notebook | ||
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| + | <div class="header-title-bottom"> | ||
| + | Lab Documents | ||
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<ul style="margin-left:20px;"> | <ul style="margin-left:20px;"> | ||
| - | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/ | + | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Overview</a></li> |
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Background</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Background</a></li> | ||
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Achievements">Achievements</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Achievements">Achievements</a></li> | ||
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length">Length Variation</a></li> | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length">Length Variation</a></li> | ||
| - | <li class="ldd_contents"><a href="">Overview</a></li> | + | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length">Overview</a></li> |
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Method">Method</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Method">Method</a></li> | ||
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Results">Results</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Results">Results</a></li> | ||
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| - | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey">High | + | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey">High School</a></li> |
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Education">Education</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Education">Education</a></li> | ||
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Survey">Survey</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Survey">Survey</a></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> |
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | pHN1257 | + | pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃ |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | KOD FX NEO | ||
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</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010 | + | R0010 |
| + | </div> | ||
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| + | each of 5.0µL DNA to DH5α | ||
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<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Cut R0010 with PstI, SpeI (using 10xH buffer) | + | Cut R0010 with PstI, SpeI (using 10xH buffer) |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | Cut pHN1257 with XbaI, PstI (using 10xM buffer) | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ligation products | + | Ligation products |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | each of 5.0µL DNA to DH5α | ||
</div> | </div> | ||
</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem- | + | R0010-stem-B0015 with 100up_F pHN1257terminator_R |
</div> | </div> | ||
</div> | </div> | ||
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<div class="step-tile"> | <div class="step-tile"> | ||
<h2 class="step-title"> | <h2 class="step-title"> | ||
| - | + | Mini-Prep | |
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<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem- | + | R0010-stem-B0015 |
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</div> | </div> | ||
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem-dT | + | R0010-stem-dT with 100up_F pHN1257terminator_R |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | KOD FX NEO | ||
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Latest revision as of 15:26, 9 September 2015
Notebook
Lab Documents
Stem Region Replacement
-
Start
-
PCR
pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃KOD FX NEO -
Transformation
R0010each of 5.0µL DNA to DH5α -
Digestion
Cut R0010 with PstI, SpeI (using 10xH buffer)Cut pHN1257 with XbaI, PstI (using 10xM buffer) -
Ligation
Ligate R0010 with pHN1257 -
Transformation
Ligation productseach of 5.0µL DNA to DH5α -
Colony PCR
R0010-stem-B0015 with 100up_F pHN1257terminator_R -
Mini-Prep
R0010-stem-B0015 -
PCR(confirm)
R0010-stem-dT with 100up_F pHN1257terminator_RKOD FX NEO -
Complete!
