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Team:HokkaidoU Japan/Notebook/Pre experiment/mRFP-expression-construct
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Latest revision as of 15:24, 9 September 2015
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!
