Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0034 mRFP Assay
From 2014.igem.org
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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 | 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 | ||
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Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) | Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) | ||
pHN1257_up_F Tm:60.1 | pHN1257_up_F Tm:60.1 | ||
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Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) | Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) | ||
LB 2 mL | LB 2 mL | ||
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LB 2 mL | LB 2 mL | ||
IPTG 20 µ or not | IPTG 20 µ or not | ||
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fluoresence imageing 514 nm 610 filter | fluoresence imageing 514 nm 610 filter | ||
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Revision as of 10:54, 14 October 2014
140910 asB0034&B0034 mRFP Assay
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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 2minColony PCR
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) pHN1257_up_F Tm:60.1 asB0034-R Tm:68.7Preculture
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) LB 2 mL for 3hours Turbidity was controlled 0.1Culture
LB 2 mL IPTG 20 µ or not for 11 hours, 13hours, 15hours, 17 hoursAssay
fluoresence imageing 514 nm 610 filterComplete!