Team:USTC CHINA/Notebook/Protocols/Extracting plasmids from gram-positive bacterium

From 2013.igem.org

(Difference between revisions)
 
(One intermediate revision not shown)
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="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" />
 
-
        </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>
<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">Home</a></li>
-
<ul id="nav">
+
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a>
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA" target="_blank">Home</a></li>
+
            <ul class="subs">
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project" target="_blank">Project</a>
+
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></li>
-
                    <ul class="subs">
+
                <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/Overview" target="_blank">Overview</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/ProjectDetails" target="_blank">Project Details</a></li>
+
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></li>
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results" target="_blank">Results</a></li>
+
            </ul>
-
                    </ul>
+
        </li>
-
                </li>
+
        <li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a>
-
                <li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook" target="_blank">Notebook</a>
+
              <ul class="subs">
-
                    <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/Timeline" target="_blank">Timeline</a></li>
+
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li>
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols" target="_blank">Protocols</a></li>
+
              </ul>
-
                    </ul>
+
        </li>
-
                </li>
+
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a>
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/" target="_blank">Modeling</a>
+
              <ul class="subs">
-
                    <ul class="subs">
+
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li>
-
                        <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">B.Subtilis Culture</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/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li>
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/MiceModeling" target="_blank">Mice Modeling</a></li>
+
              </ul>
-
                    </ul>
+
        </li>
-
              </li>
+
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a>
-
              <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice" target="_blank">Human Practice</a>
+
              <ul class="subs">
-
                    <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/Communication" target="_blank">Communication</a></li>
+
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li>
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity" target="_blank">Activity</a></li>
+
              </ul>
-
                    </ul>
+
        </li>
-
              </li>
+
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a>
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team" target="_blank">Team</a>
+
              <ul class="subs">
-
                <ul class="subs">
+
                  <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li>
-
                        <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">Profile</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/Attributions">Attributions</a></li>
-
                        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team/Attribute" target="_blank">Attribute</a></li>
+
                  <li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li>
-
                  </ul>
+
              </ul>
-
              </li>
+
        </li>
-
                <li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li>
+
        <li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li>
-
            </ul>
+
    </ul>
 +
        <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>
         </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/Extracting plasmids from gram-positive ">protocols Extracting plasmids from gram-positive </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/Extracting plasmids from gram-positive ">Extracting plasmids from gram-positive </a></div></div>
<div class="conbar2">
<div class="conbar2">
<div class="leftbar" align="left">
<div class="leftbar" align="left">
Line 71: Line 72:
<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>

Latest revision as of 09:00, 27 September 2013

Extracting plasmids from gram-positive

1.Centrifuge 2-3ml bacteria liquid with 8000rpm and remove the supernatant; resuspense the liquid with lysyme(10mg/ml)~200ul and then leave it at temperature 37℃ for 0.5-1 hour.
2.Centrifuge the liquid with 8000rpm and remove the supernatant; resuspense it with 250uL buffer P1.
3.Add 250uL buffer P2 (Alkaline lysis buffer), make it clear (reverse the tube evenly for several times) and leave it at room temperature for 2-4 minutes.
4.Add 350uL buffer P3 and reverse the tube evenly until it layers.
5.Centrifuge the tube for 10 minutes and extract plasmid extraction column from the upper water phase.
6.The following steps are identical with those of Escherichia coli.