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

From 2014.igem.org

(Difference between revisions)
 
(19 intermediate revisions not shown)
Line 1: Line 1:
{{Team:HokkaidoU_Japan/CSS}}
{{Team:HokkaidoU_Japan/CSS}}
 +
{{Team:HokkaidoU_Japan/Top_Header/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 16: 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 50: 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 83: Line 107:
</ul>
</ul>
     </div>
     </div>
-
</div><!-- end box-->
 
</div>
</div>
-
<div id="header-bottom" style="background-color:#25928E;">
+
<!--end box-->
-
  <div id="header-bottom-contents">
+
<!-- end menu-->
-
  </div>
+
-
</div>
+
-
<!--end header-->
+
<div class="wrapper">
<div class="wrapper">
Line 105: Line 125:
  <div class="step-tile">
  <div class="step-tile">
  <h2 class="step-title">
  <h2 class="step-title">
-
Transformation
+
Double Transformation
</h2>
</h2>
  </div>
  </div>
Line 118: Line 138:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-mRFP-dt on pSB6A1 on ice 30min each of 2.5&micro;L DNA to DH5&alpha; 42C    30sec on ice  2min
+
Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1)  
 +
</div>
 +
<div class="step-recipe">
 +
each of 2.5&micro;L to DH5&alpha;
</div>
</div>
    </div>
    </div>
Line 139: Line 162:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1)
+
Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1) with pHN1257_up_F Tm:60.1&#8451; & asB0034_R Tm:68.7&#8451;
-
pHN1257_up_F Tm:60.1
+
 
-
asB0034-R Tm:68.7
+
</div>
</div>
    </div>
    </div>
Line 162: Line 184:
      </ul>
      </ul>
      <div class="step-recipe">
      <div class="step-recipe">
-
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1)
+
Ptrc-stem-asB0034-stem(pHN1257) & R0040-B0034-E1010-B0015(pSB6A1)
 +
</div>
 +
<div class="step-recipe">
LB 2 mL
LB 2 mL
 +
</div>
 +
<div class="step-recipe">
for 3hours
for 3hours
 +
</div>
 +
<div class="step-recipe">
Turbidity was controlled 0.1
Turbidity was controlled 0.1
</div>
</div>
Line 187: Line 215:
      <div class="step-recipe">
      <div class="step-recipe">
LB 2 mL
LB 2 mL
 +
</div>
 +
<div class="step-recipe">
IPTG 20 &micro; or not
IPTG 20 &micro; or not
 +
</div>
 +
<div class="step-recipe">
for 11 hours, 13hours, 15hours, 17 hours
for 11 hours, 13hours, 15hours, 17 hours
</div>
</div>
Line 218: Line 250:
     </li>
     </li>
   </ul>
   </ul>
 +
</div>
</div>
</div>
</div>
</div>
<!--begin footer-->
<!--begin footer-->
-
<div id="footer-wrapper" style="background-color:#25928E;">
+
 
-
   <div id="footer-contents">
+
<div id="footer-wrapper" style="background-color: #d92424;">
-
    <div id="footer-logo">
+
   <div id="footer-content">
-
    </div>
+
<div id="footer-logo">
-
    <div id="footer-twitter">
+
<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>
 +
<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/CSS}}
+
{{Team:HokkaidoU_Japan/JS}}

Latest revision as of 15:31, 9 September 2015

Notebook
Lab Documents

140910 asB0034&B0034 mRFP Assay

  • Start

  • Double Transformation

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1)
    each of 2.5µL to DH5α

  • Colony PCR

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1) with pHN1257_up_F Tm:60.1℃ & asB0034_R Tm:68.7℃

  • Preculture

    Ptrc-stem-asB0034-stem(pHN1257) & R0040-B0034-E1010-B0015(pSB6A1)
    LB 2 mL
    for 3hours
    Turbidity was controlled 0.1

  • Culture

    LB 2 mL
    IPTG 20 µ or not
    for 11 hours, 13hours, 15hours, 17 hours

  • Assay

    fluoresence imageing 514 nm 610 filter

  • Complete!

Retrieved from "http://2014.igem.org/Team:HokkaidoU_Japan/Notebook/asB0034/140910_asB0034%26B0034_mRFP_Assay"