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Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0034 mRFP Assay
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Revision as of 10:59, 14 October 2014
140910 asB0034&B0034 mRFP Assay
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Start
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Transformation
Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42C 30sec on ice 2min -
Colony PCR
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) pHN1257_up_F Tm:60.1 asB0034-R Tm:68.7 -
Preculture
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) LB 2 mL for 3hours Turbidity was controlled 0.1 -
Culture
LB 2 mL IPTG 20 µ or not for 11 hours, 13hours, 15hours, 17 hours -
Assay
fluoresence imageing 514 nm 610 filter -
Complete!
