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Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0032 mRFP Assay
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<div class="step-tile"> | <div class="step-tile"> | ||
<h2 class="step-title"> | <h2 class="step-title"> | ||
| - | Transformation | + | Double Transformation |
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<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ptrc-stem-asB0034-stem on pHN1257& | + | Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-dt(on pSB6A1) |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | each of 2.5µL DNA to DH5α | ||
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<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | + | R0040-B0032-E1010-B0015(on pSB6A1) EcoRI_XbaI_F Tm:70.9℃ & RBS_SpeI_R Tm:59.2℃ | |
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<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ptrc-stem-asB0034-stem(pHN1257)& | + | Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-B0015(on pSB6A1) |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | LB 2 mL for 8hours | ||
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | Turbidity was controlled 0.1 | ||
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Latest revision as of 15:32, 9 September 2015
Notebook
Lab Documents
140910 asB0034 & B0032 mRFP Assay
-
Start
-
Double Transformation
Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-dt(on pSB6A1)each of 2.5µL DNA to DH5α -
Colony PCR
R0040-B0032-E1010-B0015(on pSB6A1) EcoRI_XbaI_F Tm:70.9℃ & RBS_SpeI_R Tm:59.2℃ -
Preculture
Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-B0015(on pSB6A1)LB 2 mL for 8hoursTurbidity was controlled 0.1 -
Culture
M9ZB 2 mL IPTG 20 µ or not for 9 hours, 11 hours, 13hours -
Assay
fluoresence imageing 514 nm 610 filter -
Complete!
