Team:USTC CHINA/Notebook/Protocols/Single Digestion

From 2013.igem.org

(Difference between revisions)
Line 7: Line 7:
<div class="bar" align="center">
<div class="bar" align="center">
     <div class="container" align="left">
     <div class="container" align="left">
 +
<div id="igemlogo"><a href="https://2013.igem.org/Main_Page" target="_blank"><img src="https://static.igem.org/mediawiki/2013/2/26/2013ustcigem_IGEM_basic_Logo.png" alt="igem home page" width="50" height="40" /></a></div>
 +
 +
    <ul id="nav">
 +
        <li><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a></li>
 +
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a>
 +
            <ul class="subs">
 +
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></li>
 +
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails">Project Details</a></li>
 +
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></li>
 +
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Parts">Parts</a></li>
 +
            </ul>
 +
        </li>
 +
        <li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a>
 +
              <ul class="subs">
 +
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li>
 +
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li>
 +
              </ul>
 +
        </li>
 +
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a>
 +
              <ul class="subs">
 +
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/ReporterSystem">Reporter System</a></li>
 +
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li>
 +
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/MiceModeling">Mice Modeling</a></li>
 +
              </ul>
 +
        </li>
 +
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a>
 +
              <ul class="subs">
 +
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li>
 +
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li>
 +
              </ul>
 +
        </li>
 +
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/team2">Team</a>
 +
              <ul class="subs">
 +
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/team2">Members</a></li>
 +
                  <li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li>
 +
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team/Attribution">Attribution</a></li>
 +
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team/JudgingForm">Judging Form</a></li>
 +
              </ul>
 +
        </li>
 +
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li>
 +
    </ul>
         <div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" />
         <div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" />
         </div>
         </div>
-
<div id="igemlogo"><a href="https://2013.igem.org/Main_Page" target="_blank"><img src="https://static.igem.org/mediawiki/2013/2/26/2013ustcigem_IGEM_basic_Logo.png" alt="igem home page" width="50" height="40" /></a></div>
 
-
<ul id="nav">
 
-
       
 
-
<ul id="nav">
 
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA" target="_blank">Home</a></li>
 
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project" target="_blank">Project</a>
 
-
                    <ul class="subs">
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview" target="_blank">Overview</a></li>
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails" target="_blank">Project Details</a></li>
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results" target="_blank">Results</a></li>
 
-
                    </ul>
 
-
                </li>
 
-
                <li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook" target="_blank">Notebook</a>
 
-
                    <ul class="subs">
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline" target="_blank">Timeline</a></li>
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols" target="_blank">Protocols</a></li>
 
-
                    </ul>
 
-
                </li>
 
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/" target="_blank">Modeling</a>
 
-
                    <ul class="subs">
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/ReporterSystem" target="_blank">Reporter System</a></li>
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture" target="_blank">B.Subtilis Culture</a></li>
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/MiceModeling" target="_blank">Mice Modeling</a></li>
 
-
                    </ul>
 
-
              </li>
 
-
              <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice" target="_blank">Human Practice</a>
 
-
                    <ul class="subs">
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" target="_blank">Communication</a></li>
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity" target="_blank">Activity</a></li>
 
-
                    </ul>
 
-
              </li>
 
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team" target="_blank">Team</a>
 
-
                <ul class="subs">
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team/Members" target="_blank">Members</a></li>
 
-
                        <li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA" target="_blank">Profile</a></li>
 
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team/Attribute" target="_blank">Attribute</a></li>
 
-
                  </ul>
 
-
              </li>
 
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li>
 
-
            </ul>
 
         </div>
         </div>
         </div>
         </div>
<div class="content" align="center">
<div class="content" align="center">
<div class="conbar1">
<div class="conbar1">
-
<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">notebook</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">protocols</a>&gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols/Single Digestion">single digestion</a></div></div>
+
<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a>&gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols/Single Digestion">Single Digestion</a></div></div>
<div class="conbar2">
<div class="conbar2">
<div class="leftbar" align="left">
<div class="leftbar" align="left">
Line 96: Line 97:
<div class="rightbar">
<div class="rightbar">
-
<div class="port-sidebar-border"><h>notebook</h></div>
+
<div class="port-sidebar-border"><h>Notebook</h></div>
<div class="clear"></div>
<div class="clear"></div>
-
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">timeline</a></div>
+
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></div>
-
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">protocols</a></div></div>
+
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></div></div>
</div>
</div>
</div>
</div>
</body>
</body>
</html>
</html>

Revision as of 00:30, 26 September 2013

Single Digestion

1.Performed with Thermo Scientific FastDigest. There are four restriction endonucleases we used in our experiments: EcoRI, XbaI, SpeI, PstI.
Single Digestion

Volume (μl)
Plasmid DNA Up to 1μg
10×FastDigest Buffer 2
Enzyme 1
ddH2O To 20
Total 20
2. Mix gently and spin down.
3. Incubate at 37°C in a heat block or water thermostat for 30 min.
4. Inactivate the enzyme by adding glycerol DNA loading buffer, or by heating for 5 min at 80°C(only for EcoRI) or 20min at 65°C(only for XbaI)