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Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement
From 2014.igem.org
(Difference between revisions)
| Line 131: | Line 131: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | pHN1257 | + | pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃ KOD FX NEO |
</div> | </div> | ||
</div> | </div> | ||
| Line 236: | Line 236: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem-dT | + | R0010-stem-dT 100up_F pHN1257terminator_R |
</div> | </div> | ||
</div> | </div> | ||
| Line 278: | Line 278: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem-dT | + | R0010-stem-dT 100up_F pHN1257terminator_R KOD FX NEO |
</div> | </div> | ||
</div> | </div> | ||
Revision as of 13:37, 17 October 2014
hogehoge
Stem Region Replacement
-
Start
-
PCR
pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃ KOD FX NEO -
Transformation
R0010 on ice 30min each of 5.0µL DNA to DH5α 42℃ 1min on ice 2min -
Digestion
Cut R0010 with PstI, SpeI (using 10xH buffer) and pHN1257 with XbaI, PstI (using 10xM buffer) -
Ligation
Ligate R0010 with pHN1257 -
Transformation
Ligation products on ice 30min each of 5.0µL DNA to DH5α 42℃ 1min on ice 2min -
Colony PCR
R0010-stem-dT 100up_F pHN1257terminator_R -
mini-Prep
R0010-stem-dT -
PCR(confirm)
R0010-stem-dT 100up_F pHN1257terminator_R KOD FX NEO -
Complete!
