"
Team:HokkaidoU Japan/Notebook/Pre experiment/asmRFP
From 2014.igem.org
(Difference between revisions)
| Line 128: | Line 128: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | asB0034-f Tm: 68.7& | + | asB0034-f Tm: 68.7℃ asB0034-r Tm: 68.7℃ |
</div> | </div> | ||
</div> | </div> | ||
| Line 233: | Line 233: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ptrc-stem-asB0034-stem(pHN1257) on ice 30min each of 5.0µL DNA to DH5α 42& | + | Ptrc-stem-asB0034-stem(pHN1257) on ice 30min each of 5.0µL DNA to DH5α 42℃ 1min on ice 2min |
</div> | </div> | ||
</div> | </div> | ||
| Line 254: | Line 254: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ptrc-stem-asB0034-stem(pHN1257) pHN1257_up_F Tm:60.1 asB0034-R Tm:68.7 | + | Ptrc-stem-asB0034-stem(pHN1257) pHN1257_up_F Tm:60.1℃ asB0034-R Tm:68.7℃ |
</div> | </div> | ||
</div> | </div> | ||
| Line 317: | Line 317: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α | + | Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42℃ 30sec on ice 2min |
</div> | </div> | ||
</div> | </div> | ||
Revision as of 13:00, 17 October 2014
hogehoge
anti-sense mRFP Construction
-
Start
-
Primer Annealing
asB0034-f Tm: 68.7℃ asB0034-r Tm: 68.7℃ -
Digestion
Cut pHN1257 with NcoI, XhoI(using 10×cut smart) -
Gel extraction
digested pHN1257 -
Ethanol Precipitation
digested pHN1257 -
Ligation
Ligate pHN1257 with asB0034 -
Transformation
Ptrc-stem-asB0034-stem(pHN1257) on ice 30min each of 5.0µL DNA to DH5α 42℃ 1min on ice 2min -
Colony PCR
Ptrc-stem-asB0034-stem(pHN1257) pHN1257_up_F Tm:60.1℃ asB0034-R Tm:68.7℃ -
Culture
Ptrc-stem-asB0034-stem -
mini-Prep
Ptrc-stem-asB0034-stem -
Transformation
Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42℃ 30sec on ice 2min -
Complete!
