"
Team:HokkaidoU Japan/Notebook/Pre experiment/mRFP-expression-construct
From 2014.igem.org
(Difference between revisions)
| Line 89: | Line 89: | ||
<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> |
</ul> | </ul> | ||
</div> | </div> | ||
Revision as of 02:45, 18 October 2014
Notebook
Lab Documents
mRFP Expression Plasmid Construction
-
Start
-
Get genes
pSB6A1 -
Transformation
pSB6A1 1µL to JM109 -
Liquid Culture
pSB6A1 -
Mini-prep
pSB6A1 -
PCR & PCR purification
R0040-B0034-E1010-B0015 from our stock 100up_F Tm:70.4℃ & 200dn_R Tm:66.6℃ KOD plus Neo -
Digestion
Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10×H buffer) Cut pSB6A1 with EcoRI, PstI (using 10×H buffer) -
Gel Extraction
R0040-B0034-E1010-B0015 pSB6A1 -
Ethanol precipetation
R0040-B0034-E1010-B0015 pSB6A1 -
Dephosphorylation
R0040-B0034-E1010-B0015 -
Ligation
Ligate R0040-B0034-E1010-B0015 with pSB6A1 -
Transformation
R0040-B0034-E1010-B0015(on pSB6A1) 5µL to DH5α Turbo -
Liquid Culture
R0040-B0034-E1010-B0015(on pSB6A1) -
Mini-prep
R0040-B0034-E1010-B0015(on pSB6A1) -
Complete!
