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Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement
From 2014.igem.org
(Difference between revisions)
| Line 141: | Line 141: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃ KOD FX NEO | + | pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃ |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | KOD FX NEO | ||
</div> | </div> | ||
</div> | </div> | ||
| Line 162: | Line 165: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010 | + | R0010 |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | each of 5.0µL DNA to DH5α | ||
</div> | </div> | ||
</div> | </div> | ||
| Line 183: | Line 189: | ||
</ul> | </ul> | ||
<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) | ||
</div> | </div> | ||
</div> | </div> | ||
| Line 225: | Line 234: | ||
</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> | ||
| Line 246: | Line 258: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem- | + | R0010-stem-B0015 with 100up_F pHN1257terminator_R |
</div> | </div> | ||
</div> | </div> | ||
| Line 254: | Line 266: | ||
<div class="step-tile"> | <div class="step-tile"> | ||
<h2 class="step-title"> | <h2 class="step-title"> | ||
| - | + | Mini-Prep | |
</h2> | </h2> | ||
</div> | </div> | ||
| Line 267: | Line 279: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem- | + | R0010-stem-B0015 |
</div> | </div> | ||
</div> | </div> | ||
| Line 288: | Line 300: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem-dT 100up_F pHN1257terminator_R KOD FX NEO | + | R0010-stem-dT with 100up_F pHN1257terminator_R |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | KOD FX NEO | ||
</div> | </div> | ||
</div> | </div> | ||
Revision as of 03:41, 18 October 2014
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!
