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

From 2013.igem.org

(Difference between revisions)
 
Line 9: Line 9:
<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>
         <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>
         <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a>
Line 16: Line 16:
                 <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/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/Results">Results</a></li>
-
                 <li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Parts">Parts</a></li>
+
                 <li><a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></li>
             </ul>
             </ul>
         </li>
         </li>
Line 27: Line 27:
         <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a>
         <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a>
               <ul class="subs">
               <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/KillSwitch">Kill Switch</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">B.Subtilis Culture</a></li>
-
                   <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/MiceModeling">Mice Modeling</a></li>
+
                   <li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li>
               </ul>
               </ul>
         </li>
         </li>
Line 38: Line 38:
               </ul>
               </ul>
         </li>
         </li>
-
         <li><a href="https://2013.igem.org/Team:USTC_CHINA/team2">Team</a>
+
         <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a>
               <ul class="subs">
               <ul class="subs">
-
                   <li><a href="https://2013.igem.org/Team:USTC_CHINA/team2">Members</a></li>
+
                   <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">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">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/Attributions">Attributions</a></li>
-
                   <li><a href="https://2013.igem.org/Team:USTC_CHINA/Team/JudgingForm">Judging Form</a></li>
+
                   <li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li>
               </ul>
               </ul>
         </li>
         </li>

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.

Notebook

Retrieved from "http://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols/Extracting_plasmids_from_gram-positive_bacterium"