Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0034 GFP Assay
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<h2 class="step-title"> | <h2 class="step-title"> | ||
- | Transformation | + | Double Transformation |
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- | Ptrc-stem-asB0034-stem on pHN1257& | + | Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on pSB6A1) |
- | + | </div> | |
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+ | each of 2.5µL DNA to DH5α | ||
+ | |||
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- | Ptrc-stem-asB0034-stem(pHN1257)& | + | Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(pSB6A1) pHN1257_up_F Tm:60.1℃ & asB0034-R Tm:68.7℃ |
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- | Ptrc-stem-asB0034-stem(pHN1257)& | + | Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on pSB6A1) |
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- | + | LB 2 mL for 9hours | |
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+ | Turbidity was controlled 0.1 | ||
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- | M9ZB 2 mL | + | M9ZB 2 mL IPTG 20 µ or not |
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for 9 hours, 11 hours, 13hours | for 9 hours, 11 hours, 13hours | ||
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<div class="step-recipe"> | <div class="step-recipe"> | ||
- | + | fluorescence imaging 488 nm 530 filter | |
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Latest revision as of 15:31, 9 September 2015
Notebook
Lab Documents
140910 asB0034 & B0034 GFP Assay
-
Start
-
Double Transformation
Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on pSB6A1)each of 2.5µL DNA to DH5α -
Colony PCR
Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(pSB6A1) pHN1257_up_F Tm:60.1℃ & asB0034-R Tm:68.7℃ -
Preculture
Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on pSB6A1)LB 2 mL for 9hoursTurbidity was controlled 0.1 -
Culture
M9ZB 2 mL IPTG 20 µ or notfor 9 hours, 11 hours, 13hours -
Assay
fluorescence imaging 488 nm 530 filter -
Complete!