Team:HokkaidoU Japan/Notebook/Pre experiment/Plac-failed-experiment

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Notebook

Lab Documents

Wrong Plac failed Experiment

Start
Get anti-sense vector
- Fri Jun 20
pHN1257 Great thanks to N. Nakashima
Transformation
- Thr Jul 3
R0010-B0034-E1010-B0015(pSB6A1) from destribution kit 1µL DNA to JM109
Liquid Culture
- Fri Jul 4
pSB6A1
Mini-prep
- Sat Jul 5
pSB6A1
PCR & PCR purification
- Fri Jul 25
R0040-B0034-E1010-B0015 as RBS XhoI, as mRFP NcoI
Ehanol precipetation
- Sat Jul 26
anti-sense(mRFP)
Digestion
- Sun Jul 27
Cut pHN1257 with NcoI, XhoI (using 10×Cut Smart) Cut anti-sense(mRFP) with NcoI, XhoI (using 10×Cut Smart)
Gel Extraction & Ethanol precipetation
- Sun Jul 27
pHN1257 anti-sense(mRFP)
Ligation
- Sun Jul 27
Ligate anti-sense(mFP) with pHN1257
Transformation
- Mon Jul 28
anti-sense(mRFP) on pHN1257 5µL DNA to DH5α Turbo
Colony PCR
- Tue Jul 29
anti-sense(mRFP) on pHN1257
Liquid Culture
- Wed Jul 30
anti-sense(mRFP) on pHN1257
Mini-prep
- Thr Jul 31
anti-sense(mRFP) on pHN1257
Transformation
- Sun Aug 10
anti-sense(mRFP) on pHN1257 & pSB6A1 2.5µL each DNA to DH5α Turbo
Asssay(unsuccess)
- Wed Aug 13
Culture anti-sense(mRFP) on pHN1257 & pSB6A1 1. 2mL LB with 100µL 100mM IPTG 2. 2mL LB However, IPTG was too much to grow normally.
Assay(unsuccess)
- Thr Aug 14
Culture anti-sense(mRFP) on pHN1257 & pSB6A1 E. coli was early growth 1. 2mL LB with 20µL 100mM IPTG 2. 2mL LB However, pLac was inducible promoter that is promoted by IPTG. We decided to retry this examination.
Failure...