DNA Precipitation protocol

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Revision as of 00:20, 29 September 2009 by Vaishnavi Ananth (talk | contribs) (New page: <html> <h2>Solutions/reagents:</h2><ul type="circle"><li>3M Sodium Acetate solution</li><li>Glycogen</li><li>95% EtOH</li><li>70% EtOH</li><li>water</li><li>buffer</li><li>DNA sample</li><...)
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<html> <h2>Solutions/reagents:</h2><ul type="circle"><li>3M Sodium Acetate solution</li><li>Glycogen</li><li>95% EtOH</li><li>70% EtOH</li><li>water</li><li>buffer</li><li>DNA sample</li></ul><h2>Equipment:</h2><ul type="circle"><li>Centrifuge</li><li>Eppendorf tubes</li></ul><h2>Steps:</h2><ol><p><li>Measure out DNA sample into an Eppendorf tube.<br>Add <b><font color=#357EC7>0.1</font></b> volume <font color=#357EC7>3M Sodium Acetate solution</font>.<br></li></p><p><li>Add <b><font color=#357EC7>1 µl</font></b> of <font color=#357EC7>Glycogen</font>.<br></li></p><p><li>Add <b><font color=#357EC7>2</font></b> volumes <font color=#357EC7>95% EtOH</font>.<br></li></p><p><li><p>Option 1: Store at <b><font color=#357EC7>-20°C</font></b> for <b><font color=#357EC7>12 hrs</font></b>(overnight).<br>(or)<br>Option 2: Store at <b><font color=#357EC7>-80°C</font></b> for <b><font color=#357EC7>30 mins</font></b>.<br></p><p></li></p><p><li>Centrifuge at <font color=#357EC7>maximum speed</font> for at least <b><font color=#357EC7>15 mins</font></b> at <b><font color=#357EC7>room temperature</font></b>, gently aspirate out the supernatant and discard it.<br></li></p><p><b><font size=3>(Optional)</font></b><br>Add <b><font color=#357EC7>1 ml</font></b> of <font color=#357EC7>70% EtOH</font>.<br>Store at <b><font color=#357EC7><b><font color=#357EC7>room temperature</font></b></font></b> for <b><font color=#357EC7>5 mins</font></b>.<br></li></p><p><b><font size=3>(Optional)</font></b><br>Centrifuge at <font color=#357EC7>maximum speed</font> for <b><font color=#357EC7>5 mins</font></b> at <b><font color=#357EC7>room temperature</font></b>, gently aspirate out the supernatant and discard it.<br></li></p><p><li>Dry the pellet in air for <b><font color=#357EC7>10 - 15 mins</font></b>.<br><font color = "#800517"><i>Dry until all the liquid is gone.</i></font><br></li></p><p><li><p>Option 1: Add <font color=#357EC7>water</font> to pellet.<br>(or)<br>Option 2: Add <font color=#357EC7>buffer</font> to pellet.<br></p><p>Resuspend the pellet by vortexing/by shaking vigorously.<br></li></p></ol></html>