Team:Tuebingen/Notebook/Protocols/preparative-restriction

From 2013.igem.org

(Difference between revisions)
 
(2 intermediate revisions not shown)
Line 16: Line 16:
<div id="actualContent">  
<div id="actualContent">  
-
<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a>
 
-
 
-
<p>&nbsp;</p>
 
<h3>Reagents</h3>
<h3>Reagents</h3>
Line 74: Line 71:
</ol>
</ol>
 +
 +
<p><a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a></p>
</div>
</div>

Latest revision as of 12:11, 4 October 2013

Return to iGEM Main Page.

Preparative Restriction Digest

Reagents

10.0 µL 10x NEBuffer 2
1.0 µL 100x BSA
5 - 10 µg Plasmid DNA
2.0 µL EcoRI or XbaI
2.0 µL PstI or SpeI
to 100 µL Aqua dest.
1/10 vol 10X Antarctic Phosphatase

 

Procedure

  1. Mix all reagents (except for Antarctic Phosphatase) in an 1.5 mL Eppendorf-tube and incubate at 37 °C over night.
  2. On the next day, heat inactivate restriction enzymes at 80°C for 20 min.
  3. Add 1/10 volume of 10X Antarctic Phosphatase and mix. Incubate at 37 °C for 15 min for 5' extensions.
  4. Heat inactivate phosphatase at 70 °C for 5 min.
  5. Run gel and perform a gelextraction.

Back to Protocols