Team:HokkaidoU Japan/Notebook/Pre experiment/asmRFP

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Notebook

Lab Documents

Anti-Sense mRFP Construction

Start
Get anti-sense vector
- Fri Jun 20
Great thanks to N. Nakashima
PCR & PCR purification
- Fri Aug 15
R0040-B0034-E1010-B0015 with as_RBS_XhoI_F Tm:68.2℃ & as_mRFP_NcoI_R Tm:76.0℃

KOD plus Neo
Ethanol Precipitation
- Fri Aug 15
asmRFP
Digestion
- Fri Aug 15
Cut pHN1257 with NcoI, XhoI (using 10×Cut Smart)

Cut asmRFP with NcoI, XhoI (using 10×Cut Smart)
Gel Extraction
- Sat Aug 16
pHN1257 asmRFP
Ethanol Precipitation
- Sat Aug 16
asmRFP
Dephosphorylation
- Sat Aug 16
asmRFP
Ligation
- Sat Aug 16
Ligate asmRFP with pHN1257
Transformation
- Sat Aug 16
asmRFP(on pHN1257)

5µL to DH5α Turbo
Colony PCR
- Sun Aug 17, Mon Aug 18
asmRFP(on pHN1257) pHN1257_up_F Tm:60.1℃ & pHN1257_terminator_R Tm:71.4℃

Kapa-Taq
Liquid Culture
- Mon Aug 18
asmRFP(on pHN1257)
Mini-Prep
- Tue Aug 19
asmRFP(pHN1257)
Complete!