Team:HokkaidoU Japan/Notebook/Pre experiment/mRFP-expression-construct

From 2014.igem.org

(Difference between revisions)
 
(12 intermediate revisions not shown)
Line 1: Line 1:
{{Team:HokkaidoU_Japan/CSS}}
{{Team:HokkaidoU_Japan/CSS}}
-
{{Team:HokkaidoU_Japan/Header/CSS}}
+
{{Team:HokkaidoU_Japan/Top_Header/CSS}}
{{Team:HokkaidoU_Japan/Footer/CSS}}
{{Team:HokkaidoU_Japan/Footer/CSS}}
{{Team:HokkaidoU_Japan/Notebook/CSS}}
{{Team:HokkaidoU_Japan/Notebook/CSS}}
-
{{Team:HokkaidoU_Japan/LargeDropDown_Projects/CSS}}
+
{{Team:HokkaidoU_Japan/LargeDropDown/CSS}}
<html>
<html>
 +
<link href='http://fonts.googleapis.com/css?family=Merriweather' rel='stylesheet' type='text/css'>
<div id="background">
<div id="background">
<!--begin header-->
<!--begin header-->
-
<div id="header-top" style="background-color:#6CC4C1;">
+
<div id="header-top">
 +
<img src="https://static.igem.org/mediawiki/2014/b/bb/HokkaidoU_Header_Top.png">
 +
</div>
 +
<div id="header-bottom">
 +
  <div id="header-bottom-contents">
 +
    <div class="header-title">
 +
      <div class="header-title-top">
 +
      Notebook
 +
      </div>
 +
      <div class="header-title-bottom">
 +
      Lab Documents
 +
      </div>
 +
    </div>
 +
    <div class="header-picture">
 +
      <img src="https://static.igem.org/mediawiki/2014/4/44/HokkaidoU_Header_ki.png">
 +
    </div>
 +
  </div>
 +
</div>
 +
<!--end header-->
 +
<!--begin menu-->
     <div class="box">
     <div class="box">
   <div id="box-wrapper">  
   <div id="box-wrapper">  
 +
       <ul id="ldd_menu" class="ldd_menu">
       <ul id="ldd_menu" class="ldd_menu">
                                 <li>
                                 <li>
-
                                 <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-->
+
                                 <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-->
                                 </li>
                                 </li>
<li class="menu_button">
<li class="menu_button">
Line 18: Line 39:
<div class="ldd_submenu" style="left:-415px; width:970px;">
<div class="ldd_submenu" style="left:-415px; width:970px;">
<ul style="margin-left:20px;">
<ul style="margin-left:20px;">
-
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Introduction">Overview</a></li>
+
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Overview</a></li>
-
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Introduction">Introduction</a></li>
+
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Background</a></li>
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Background</a></li>
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Achievements">Achievements</a></li>
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Achievements">Achievements</a></li>
</ul>
</ul>
<ul>
<ul>
-
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem">H-Stem System</a></li>
+
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem">H-stem System</a></li>
                                                         <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#Overview">Overview</a></li>
                                                         <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#Overview">Overview</a></li>
 +
                                                        <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#asRNA_stabilization">Stabilization</a></li>
 +
                                                        <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#Modelling">Modelling</a></li>
 +
                                                        <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#Construct">Construct</a></li>
                                                         <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#How_To_Use">How To Use</a></li>
                                                         <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/H_Stem#How_To_Use">How To Use</a></li>
-
                                                        <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Project/H_Stem#Modeling">Modelling</a></li>
 
</ul>
</ul>
<ul>
<ul>
-
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Overview">Anti-senseB0034</a></li>
+
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034">Anti-sense B0034</a></li>
-
            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Overview">Overview</a></li>
+
            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034">Overview</a></li>
-
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Method">Method</a></li>
+
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034#Method">Method</a></li>
-
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Results">Results</a></li>
+
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034#Results">Results</a></li>
-
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034/Conclusion">Conclusion</a></li>  
+
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/asB0034#Conclusion">Conclusion</a></li>
-
                                                </ul>
+
</ul>
<ul>
<ul>
-
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Overview">Length Variation</a></li>
+
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length">Length Variation</a></li>
-
            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Overview">Overview</a></li>
+
            <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length">Overview</a></li>
-
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Method">Method</a></li>
+
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Method">Method</a></li>
-
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Results">Results</a></li>
+
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Results">Results</a></li>
-
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length/Conclusiton">Conclusion</a></li>
+
                                                             <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Conclusion">Conclusion</a></li>
</ul>
</ul>
<ul>
<ul>
Line 52: Line 74:
</li>
</li>
<li class="menu_button">
<li class="menu_button">
-
<span class="menu-list">Policy and Practice</span>
+
<span class="menu-list"><span style="position:relative; top:10px;">Outreach</span></span>
<div class="ldd_submenu" style="width:590px; left:-230px;">
<div class="ldd_submenu" style="width:590px; left:-230px;">
<ul style="margin-left:20px;">
<ul style="margin-left:20px;">
-
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Education">Education</a></li>
+
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Univ_festival">Univ. Festival</a></li>
-
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Education#School_Festival">School Festival</a></li>
+
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Univ_festival#Education">Education</a></li>
-
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Education#High_School">High-school</a></li>
+
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Univ_festival#Survey">Survey</a></li>
</ul>
</ul>
<ul>
<ul>
-
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Survey">Survey</a></li>
+
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey">High School</a></li>
-
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Survey#School_Festival">School Festival</a></li>
+
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Education">Education</a></li>
-
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/Survey#High_School">High-school</a></li>
+
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Survey">Survey</a></li>
</ul>
</ul>
<ul>
<ul>
-
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Policy_And_Practice/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/Policy_And_Practice/Discussion#Background">Background</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/Policy_And_Practice/Discussion#Estimation">Estimation</a></li>
+
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Evaluation">Evaluation</a></li>
</ul>
</ul>
</div>
</div>
Line 85: Line 107:
</ul>
</ul>
     </div>
     </div>
-
</div><!-- end box-->
 
</div>
</div>
-
<div id="header-bottom" style="background-color:#25928E;">
+
 
-
  <div id="header-bottom-contents">
+
<!--end box-->
-
  </div>
+
<!-- end menu-->
-
</div>
+
-
<!--end header-->
+
<div class="wrapper">
<div class="wrapper">
   <div id="hokkaidou-contents">
   <div id="hokkaidou-contents">
Line 128: Line 147:
      </ul>
      </ul>
      <div class="step-recipe"> pSB6A1
      <div class="step-recipe"> pSB6A1
-
1&micro;L DNA to JM109</div>
+
1&micro;L to JM109</div>
    </div>
    </div>
  </div>
  </div>
Line 174: Line 193:
      </ul>
      </ul>
      <div class="step-recipe"> R0040-B0034-E1010-B0015 from our stock
      <div class="step-recipe"> R0040-B0034-E1010-B0015 from our stock
-
100up 200dn</div>
+
100up_F Tm:70.4&#8451; & 200dn_R Tm:66.6&#8451;
 +
</div>
 +
<div class="step-recipe">
 +
        KOD plus Neo</div>
    </div>
    </div>
  </div>
  </div>
Line 190: Line 212:
      </ul>
      </ul>
      <div class="step-recipe"> Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10&times;H buffer)
      <div class="step-recipe"> Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10&times;H buffer)
-
Cut pSB6A1 with EcoRI, PstI (using 10&times;H buffer)</div>
+
</div>
 +
<div class="step-recipe">
 +
Cut pSB6A1 with EcoRI, PstI (using 10&times;H buffer)
 +
</div>
    </div>
    </div>
  </div>
  </div>
Line 196: Line 221:
<li class="step">
<li class="step">
  <div class="step-tile">
  <div class="step-tile">
-
  <h2 class="step-title">Gel Extraction & Ethanol precipetation</h2>
+
  <h2 class="step-title">Gel Extraction</h2>
  </div>
  </div>
  <div class="step-detail">
  <div class="step-detail">
Line 206: Line 231:
      </ul>
      </ul>
      <div class="step-recipe"> R0040-B0034-E1010-B0015
      <div class="step-recipe"> R0040-B0034-E1010-B0015
-
pSB6A1</div>
+
pSB6A1
 +
</div>
 +
    </div>
 +
  </div>
 +
</li>
 +
<li class="step">
 +
  <div class="step-tile">
 +
  <h2 class="step-title">Ethanol precipetation</h2>
 +
  </div>
 +
  <div class="step-detail">
 +
    <div class="step-detail-body">
 +
      <ul class="step-detail-trials">
 +
        <li>
 +
  <time>Sat Aug 16</time>
 +
</li>
 +
      </ul>
 +
      <div class="step-recipe">R0040-B0034-E1010-B0015
 +
pSB6A1
 +
</div>
    </div>
    </div>
  </div>
  </div>
Line 221: Line 264:
</li>
</li>
      </ul>
      </ul>
-
      <div class="step-recipe"> R0040-B0034-E1010-B0015</div>
+
      <div class="step-recipe"> R0040-B0034-E1010-B0015
 +
</div>
    </div>
    </div>
  </div>
  </div>
Line 236: Line 280:
</li>
</li>
      </ul>
      </ul>
-
      <div class="step-recipe"> Ligate R0040-B0034-E1010-B0015 with pSB6A1</div>
+
      <div class="step-recipe"> Ligate R0040-B0034-E1010-B0015 with pSB6A1
 +
</div>
    </div>
    </div>
  </div>
  </div>
Line 251: Line 296:
</li>
</li>
      </ul>
      </ul>
-
      <div class="step-recipe"> R0040-B0034-E1010-B0015 on pSB6A1
+
      <div class="step-recipe"> R0040-B0034-E1010-B0015(on pSB6A1)
-
5&micro;L DNA to DH5&alpha; Turbo</div>
+
5&micro;L to DH5&alpha; Turbo
 +
</div>
    </div>
    </div>
  </div>
  </div>
Line 267: Line 313:
</li>
</li>
      </ul>
      </ul>
-
      <div class="step-recipe"> R0040-B0034-E1010-B0015 on pSB6A1</div>
+
      <div class="step-recipe"> R0040-B0034-E1010-B0015(on pSB6A1)
 +
</div>
    </div>
    </div>
  </div>
  </div>
Line 282: Line 329:
</li>
</li>
      </ul>
      </ul>
-
      <div class="step-recipe"> R0040-B0034-E1010-B0015 on pSB6A1</div>
+
      <div class="step-recipe">R0040-B0034-E1010-B0015(on pSB6A1)</div>
    </div>
    </div>
  </div>
  </div>
Line 293: Line 340:
</div>
</div>
</div>
</div>
-
<div id="footer-wrapper" style="background-color:#25928E;">
+
<!--begin footer-->
-
   <div id="footer-contents">
+
 
-
    <div id="footer-logo">
+
<div id="footer-wrapper" style="background-color: #d92424;">
-
    </div>
+
   <div id="footer-content">
-
    <div id="footer-twitter">
+
<div id="footer-logo">
-
    </div>
+
<a href="http://igemhokkaidou.wordpress.com"><img style="height:150px;position:relative;" src="https://static.igem.org/mediawiki/2014/3/39/HokkaidoU_logo_transparent.png"></a>
-
  </div>
+
</div>
</div>
 +
<div id="footer-twitter">
 +
        <a href="http://twitter.com/igem_hokkaidou"><img style="height:150px; position:relative; bottom:0;" src="https://static.igem.org/mediawiki/2014/3/3b/HokkaidoU_Footer_Twitter.png"></a>
 +
    </div>
-
<!--end footer-->
+
  </div>
 +
</div>
</div><!--background-->
</div><!--background-->
 +
<link href='http://fonts.googleapis.com/css?family=Special+Elite' rel='stylesheet' type='text/css'>
</html>
</html>
{{Team:HokkaidoU_Japan/JS}}
{{Team:HokkaidoU_Japan/JS}}

Latest revision as of 15:24, 9 September 2015

Notebook
Lab Documents

mRFP Expression Plasmid Construction

  • Start

  • Get genes

    pSB6A1

  • Transformation

    pSB6A1 1µL to JM109

  • Liquid Culture

    pSB6A1

  • Mini-prep

    pSB6A1

  • PCR & PCR purification

    R0040-B0034-E1010-B0015 from our stock 100up_F Tm:70.4℃ & 200dn_R Tm:66.6℃
    KOD plus Neo

  • Digestion

    Cut R0040-B0034-E1010-B0015 with EcoRI, PstI (using 10×H buffer)
    Cut pSB6A1 with EcoRI, PstI (using 10×H buffer)

  • Gel Extraction

    R0040-B0034-E1010-B0015 pSB6A1

  • Ethanol precipetation

    R0040-B0034-E1010-B0015 pSB6A1

  • Dephosphorylation

    R0040-B0034-E1010-B0015

  • Ligation

    Ligate R0040-B0034-E1010-B0015 with pSB6A1

  • Transformation

    R0040-B0034-E1010-B0015(on pSB6A1) 5µL to DH5α Turbo

  • Liquid Culture

    R0040-B0034-E1010-B0015(on pSB6A1)

  • Mini-prep

    R0040-B0034-E1010-B0015(on pSB6A1)

  • Complete!

Retrieved from "http://2014.igem.org/Team:HokkaidoU_Japan/Notebook/Pre_experiment/mRFP-expression-construct"