Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement

From 2014.igem.org

(Difference between revisions)
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>
 +
<div id="background">
<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">
 +
hogehoge
 +
  </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 29:
<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/Introduction">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/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>
Line 24: Line 35:
</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="">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 65:
</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#Estimation">Evaluation</a></li>
</ul>
</ul>
</div>
</div>
Line 85: Line 98:
</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">

Revision as of 13:35, 16 October 2014

hogehoge

Stem Region Replacement

  • Start

  • PCR

    pHN1257 161 Tm: 162 Tm: KOD FX NEO

  • Transformation

    R0010 on ice 30min each of 5.0µL DNA to DH5α 42°C 1min on ice 2min

  • Digestion

    Cut R0010 with PstI, SpeI (using 10xH buffer) and pHN1257 with XbaI, PstI (using 10xM buffer)

  • Ligation

    Ligate R0010 with pHN1257

  • Transformation

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

  • Colony PCR

    R0010-stem-dT 100up f 162primer R(あとで書く)

  • mini-Prep

    R0010-stem-dT

  • PCR(confirm)

    R0010-stem-dT 100up 162 KOD FX NEO

  • Complete!

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