Team:HokkaidoU Japan/Notebook/Pre experiment/asmRFP

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                                <span class="menu-list"><a href="https://2014.igem.org/Team:HokkaidoU_Japan"><span style="position:relative; top:10px;">Top</span></a></span><!-- Increases to 510px in width-->
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<ul style="margin-left:20px;">
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Introduction">Overview</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Introduction">Introduction</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Background</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Achievements">Achievements</a></li>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem">H-Stem System</a></li>
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                                                        <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#Overview">Overview</a></li>
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                                                        <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#How_To_Use">How To Use</a></li>
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                                                        <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Project/H_Stem#Modeling">Modelling</a></li>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Overview">Anti-senseB0034</a></li>
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            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Overview">Overview</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Method">Method</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Results">Results</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Conclusion">Conclusion</a></li>
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                                                </ul>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Overview">Length Variation</a></li>
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            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Overview">Overview</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Method">Method</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Results">Results</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Conclusiton">Conclusion</a></li>
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</ul>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Materials">Extra Materials</a></li>
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            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Parts">Parts</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Notebook">Notebook</a></li>
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                                                            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Safety">Safety</a></li>
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<span class="menu-list">Policy and Practice</span>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Education">Education</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Education#School_Festival">School Festival</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Education#High_School">High-school</a></li>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Survey">Survey</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Survey#School_Festival">School Festival</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Survey#High_School">High-school</a></li>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Discussion">Discussion</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Discussion#Background">Background</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Discussion#Estimation">Estimation</a></li>
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<span class="menu-list"><span style="position:relative; top:10px;">About Us</span></span>
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<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/About_Us">About Us</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/About_Us#Acknowledgements">Acknowledgements</a></li>
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<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/About_Us#Team">Team</a></li>
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{{Team:HokkaidoU_Japan/JS}}

Revision as of 11:14, 14 October 2014

anti-sense mRFP Construction

  • Start

  • Primer Annealing

    asB0034-f Tm: 68.7°C asB0034-r Tm: 68.7°C

  • 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) on ice 30min each of 5.0µL DNA to DH5α 42°C 1min on ice 2min

  • Colony PCR

    Ptrc-stem-asB0034-stem(pHN1257) 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&Ptet-B0034-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42C 30sec on ice 2min

  • Complete!

Retrieved from "http://2014.igem.org/Team:HokkaidoU_Japan/Notebook/Pre_experiment/asmRFP"