Team:HokkaidoU Japan/Notebook/asB0034/asB0034,32-1C3-construction

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Notebook

Lab Documents

Anti-Sense B0034,B0032 on 1C3 Construction

Start
Digestion
- Mon Sep 29
Cut vector with EcoRI, PstI (using 10×H buffer)

Cut insert with EcoRI, PstI (using 10×M buffer)
Digestion
- Tue Sep 30
Cut vector with EcoRI, PstI (using 10×H buffer)
Gel extraction
- Tue Sep 30
digested vector insert
Ethanol Precipitation
- Tue Sep 30
digested vector insert
Ligation
- Tue Sep 30
Ligate vector with insert
Transformation
- Tue Sep 30
ligation products

5.0µL to DH5α
Colony PCR
- Wed Oct 1
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
- Wed Oct 1
R0010-stem-asB0034-stem-B0015(on pSB1C3)

R0010-stem-asB0032-stem-B0015(on pSB1C3)
Mini-Prep
- Wed Oct 1
R0010-stem-asB0034-stem-B0015(on pSB1C3)

R0010-stem-asB0032-stem-B0015(on pSB1C3)
Sequencing
- Wed Oct 1
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!