Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0034 GFP Assay

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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion">Discussion</a></li>
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion">Discussion</a></li>
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Background">Background</a></li>
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Background">Background</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Evaluation">Evaluation</a></li>
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Transformation
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Double Transformation
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      </ul>
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      <div class="step-recipe">
      <div class="step-recipe">
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Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice  30min
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on pSB6A1)
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each of 2.5&micro;L DNA to DH5&alpha; 42&#8451;    30sec
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</div>
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on ice  2min
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<div class="step-reipe">
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    each of 2.5&micro;L DNA to DH5&alpha;
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Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1)
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(pSB6A1) pHN1257_up_F Tm:60.1&#8451; & asB0034-R Tm:68.7&#8451;
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pHN1257_up_F Tm:60.1&#8451;
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asB0034-R Tm:68.7&#8451;
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Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-GFP-dT(pSB6A1)
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-B0040-B0015(on pSB6A1)
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LB 2 mL
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</div>
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for 9hours
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<div class="step-recipe">
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Turbidity was controlled 0.1
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LB 2 mL for 9hours
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Turbidity was controlled 0.1
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      </ul>
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M9ZB 2 mL
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M9ZB 2 mL IPTG 20 &micro; or not
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IPTG 20 &micro; or not
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for 9 hours, 11 hours, 13hours
for 9 hours, 11 hours, 13hours
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      </ul>
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fluoresence imageing 488 nm 530 filter
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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 9hours
    Turbidity was controlled 0.1

  • Culture

    M9ZB 2 mL IPTG 20 µ or not
    for 9 hours, 11 hours, 13hours

  • Assay

    fluorescence imaging 488 nm 530 filter

  • Complete!

Retrieved from "http://2014.igem.org/Team:HokkaidoU_Japan/Notebook/asB0034/140910_asB0034%26B0034_GFP_Assay"