Team:USTC CHINA/Notebook/Protocols/Single Digestion

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 08:55, 27 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)

Notebook

Retrieved from "http://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols/Single_Digestion"