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Team:HokkaidoU Japan/Notebook/asB0034/asB0034,32-1C3-construction
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Latest revision as of 15:30, 9 September 2015
Notebook
Lab Documents
Anti-Sense B0034,B0032 on 1C3 Construction
-
Start
-
Digestion
Cut vector with EcoRI, PstI (using 10×H buffer)Cut insert with EcoRI, PstI (using 10×M buffer) -
Digestion
Cut vector with EcoRI, PstI (using 10×H buffer) -
Gel extraction
digested vector insert -
Ethanol Precipitation
digested vector insert -
Ligation
Ligate vector with insert -
Transformation
ligation products5.0µL to DH5α -
Colony PCR
R0010-stem-asB0034-stem-B0015(on pSB1C3) 100up_F Tm:62.6℃ & asB0034_R Tm:68.7℃R0010-stem-asB0032-stem-B0015(on pSB1C3) 100up_F Tm:62.6℃ & asB0032_R Tm:70.8℃ -
Culture
R0010-stem-asB0034-stem-B0015(on pSB1C3)R0010-stem-asB0032-stem-B0015(on pSB1C3) -
Mini-Prep
R0010-stem-asB0034-stem-B0015(on pSB1C3)R0010-stem-asB0032-stem-B0015(on pSB1C3) -
Sequencing
R0010-stem-asB0034-stem-B0015(on pSB1C3) with 100up_F Tm:62.8℃ & 200dn_R Tm:66.6℃R0010-stem-asB0032-stem-B0015(on pSB1C3) with 100up_F Tm:62.6℃ & 200dn_R Tm:66.6℃ -
Complete!
