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Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement
From 2014.igem.org
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<ul> | <ul> | ||
<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> | ||
Revision as of 21:31, 17 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
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!
