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Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0032 mRFP Assay
From 2014.igem.org
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| - | Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0032-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α | + | Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0032-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42℃ 30sec on ice 2min |
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| - | Ptet-B0032-mRFP-dT on pSB6A1 | + | Ptet-B0032-mRFP-dT on pSB6A1 EcoI_XbaI_F Tm:70.9℃ rbs_SpeI_R Tm:59.2℃ |
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Revision as of 14:45, 17 October 2014
hogehoge
140910 asB0034 & B0032 mRFP Assay
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Start
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Transformation
Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0032-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42℃ 30sec on ice 2min -
Colony PCR
Ptet-B0032-mRFP-dT on pSB6A1 EcoI_XbaI_F Tm:70.9℃ rbs_SpeI_R Tm:59.2℃ -
Preculture
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0032-mRFP-dT(pSB6A1) LB 2 mL for 8hours -
Culture
M9ZB 2 mL IPTG 20 µ or not for 9 hours, 11 hours, 13hours -
Assay
fluoresence imageing 514 nm 610 filter -
Complete!
