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Team:HokkaidoU Japan/Notebook/asB0034/asmRFP
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| - | asB0034_F Tm: 68.7℃ asB0034_R Tm: 68.7℃ | + | asB0034_F Tm: 68.7℃ & asB0034_R Tm: 68.7℃ |
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| - | Ptrc-stem-asB0034-stem(pHN1257) | + | Ptrc-stem-asB0034-stem(pHN1257) |
| + | each of 5.0µL DNA to DH5α | ||
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| - | Ptrc-stem-asB0034-stem(pHN1257) pHN1257_up_F Tm:60.1℃ asB0034_R Tm:68.7℃ | + | Ptrc-stem-asB0034-stem(pHN1257) with pHN1257_up_F Tm:60.1℃ asB0034_R Tm:68.7℃ |
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| - | Ptrc-stem-asB0034-stem on pHN1257& | + | Ptrc-stem-asB0034-stem on pHN1257 & R0040-B0034-E1010-B0015 on pSB6A1 |
| + | each of 2.5µL to DH5α | ||
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Revision as of 02:26, 18 October 2014
Notebook
Lab Documents
anti-sense mRFP Construction
-
Start
-
Primer Annealing
asB0034_F Tm: 68.7℃ & asB0034_R Tm: 68.7℃ -
Digestion
Cut pHN1257 with NcoI, XhoI(using 10×cut smart) -
Gel extraction
digested pHN1257 -
Ethanol Precipitation
digested pHN1257 -
Ligation
Ligate pHN1257 with asB0034 -
Transformation
Ptrc-stem-asB0034-stem(pHN1257) each of 5.0µL DNA to DH5α -
Colony PCR
Ptrc-stem-asB0034-stem(pHN1257) with pHN1257_up_F Tm:60.1℃ asB0034_R Tm:68.7℃ -
Culture
Ptrc-stem-asB0034-stem -
mini-Prep
Ptrc-stem-asB0034-stem -
Transformation
Ptrc-stem-asB0034-stem on pHN1257 & R0040-B0034-E1010-B0015 on pSB6A1 each of 2.5µL to DH5α -
Complete!
