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Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0034 GFP Assay
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| + | <h2>140910 asB0034 & B0034 GFP Assay</h2> | ||
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Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice 30min | Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice 30min | ||
each of 2.5µL DNA to DH5α 42C 30sec | each of 2.5µL DNA to DH5α 42C 30sec | ||
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</li> | </li> | ||
</ul> | </ul> | ||
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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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</li> | </li> | ||
</ul> | </ul> | ||
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Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-GFP-dT(pSB6A1) | Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-GFP-dT(pSB6A1) | ||
LB 2 mL | LB 2 mL | ||
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</li> | </li> | ||
</ul> | </ul> | ||
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M9ZB 2 mL | M9ZB 2 mL | ||
IPTG 20 µ or not | IPTG 20 µ or not | ||
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</li> | </li> | ||
</ul> | </ul> | ||
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fluoresence imageing 488 nm 530 filter | fluoresence imageing 488 nm 530 filter | ||
</div> | </div> | ||
Revision as of 09:08, 25 September 2014
140910 asB0034 & B0034 GFP Assay
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Start
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Transformation
- <time> Sun Sep 7 </time>
Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42C 30sec on ice 2min
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Colony PCR
- <time> Mon Sep 8 </time>
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) pHN1257_up_F Tm:60.1 asB0034-R Tm:68.7
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Preculture
- <time> Tue Sep 9 </time>
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-GFP-dT(pSB6A1) LB 2 mL for 9hours Turbidity was controlled 0.1
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Culture
- <time> Tue Sep 9 </time>
M9ZB 2 mL IPTG 20 µ or not for 9 hours, 11 hours, 13hours
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Assay
- <time> Wed Sep 10 </time>
fluoresence imageing 488 nm 530 filter
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Complete!
