Team:UNIPV-Pavia/Protocols/Precipitation

From 2009.igem.org

(Difference between revisions)
(DNA precipitation with sodium acetate)
(DNA precipitation with sodium acetate)
 
(10 intermediate revisions not shown)
Line 1: Line 1:
-
{{UNIPV-Pavia/StyleCss}}
 
<html>
<html>
-
<head>
+
 
-
<link rel="stylesheet" type="text/css" href="http://bioinfo.unipv.it/igem/templates/SpryAssets/SpryMenuBar.js" title="Stile"/>
+
-
</head>
+
-
<body>
+
<table width="100%">
<table width="100%">
   <tr>
   <tr>
<td align="center">
<td align="center">
-
<img width="100%" height="180px" src="https://static.igem.org/mediawiki/2009/c/c2/UNIPV_title_MandM.jpg" ></img>
+
<img width="100%" height="180px" src="https://static.igem.org/mediawiki/2009/d/de/UNIPV_title_M%26M.jpg" ></img>
</td>
</td>
   </tr>
   </tr>
</table>
</table>
-
</body>
+
 
</html>
</html>
Line 25: Line 21:
     <font face="Pristina" size="50" ><b>Protocols</b></font>
     <font face="Pristina" size="50" ><b>Protocols</b></font>
     </td>
     </td>
 +
    <td width="50px"></td>
</html>
</html>
   </tr>
   </tr>
Line 37: Line 34:
<br>
<br>
<table width='80%'><tr><td>
<table width='80%'><tr><td>
-
*We use it only if the miniprep did not give us a high yield of DNA
+
*Add 1/10 DNA solution volume of sodium acetate 3 M
-
*Add 1/10 digestion volume of sodium acetate 3 M
+
*Add 2.5 DNA solution volume of absolute ethanol
-
*Add 2.5 digestion volume of absolute ethanol
+
*Freeze at -80°C for 30 min
*Freeze at -80°C for 30 min
*Centrifuge at 13000 rpm, 4°C for 20 min
*Centrifuge at 13000 rpm, 4°C for 20 min
Line 47: Line 43:
*Remove all supernatant with a pipette
*Remove all supernatant with a pipette
*Air dry pellet until ethanol is totally removed
*Air dry pellet until ethanol is totally removed
-
*Elute with 5 µl ddH2O
+
*Elute with 5-10 µl of ddH2O
<br><br>
<br><br>
-
<br></td><td align='center' valign='bottom' width='300px'>
+
<br></td><td align='center' valign='top' width='300px'>
-
<div align='center'>
+
 
<table border='1'>
<table border='1'>
 +
<tr bgcolor='yellow'><td>INGREDIENTS:
 +
</td></tr>
<tr><td>
<tr><td>
-
*'''Sodium acetate'''
+
<font class='label'>
-
*'''Absolute ethanol'''
+
'''Sodium acetate'''<hr color='white' width='70%'>
-
*'''Ethanol 70%'''
+
'''Absolute ethanol'''<hr color='white' width='70%'>
-
*'''ddH2O'''
+
'''Ethanol 70%'''<hr color='white' width='70%'>
-
*'''Previously cut plasmid'''
+
'''ddH2O'''<hr color='white' width='70%'>
-
</td></tr></table></div><br><br><br><br>
+
'''DNA sample'''</font>
-
</td></tr></table
+
</td></tr></table><br><br><br><br>
 +
</td></tr></table>

Latest revision as of 10:32, 21 October 2009

EthanolPVanimation.gif



Protocols

DNA precipitation with sodium acetate

(estimated time: 1 hour)

  • Add 1/10 DNA solution volume of sodium acetate 3 M
  • Add 2.5 DNA solution volume of absolute ethanol
  • Freeze at -80°C for 30 min
  • Centrifuge at 13000 rpm, 4°C for 20 min
  • Decant supernatant
  • Add 50 µl of 70% ethanol
  • Centrifuge at 13000 rpm, 4°C for 20 min
  • Remove all supernatant with a pipette
  • Air dry pellet until ethanol is totally removed
  • Elute with 5-10 µl of ddH2O




INGREDIENTS:

Sodium acetate
Absolute ethanol
Ethanol 70%
ddH2O

DNA sample