DNA Precipitation protocol: Difference between revisions
(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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<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 | <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></ul><h2>Steps:</h2><ol><p><li>Measure out <b><font color=#357EC7>0.1</font></b> volume <font color=#357EC7>3M Sodium Acetate solution</font> into DNA sample.<br></li></p><p><li>Measure out <b><font color=#357EC7>1 µl</font></b> of <font color=#357EC7>Glycogen</font> into Eppendorf tube (1).<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><b>Option 1: </b>Store at <b><font color=#357EC7>-20°C</font></b> for <b><font color=#357EC7>12 hrs</font></b>(overnight).<br>(or)<br><b>Option 2: </b>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><b>Option 1: </b>Add <font color=#357EC7>water</font> to pellet.<br>(or)<br><b>Option 2: </b>Add <font color=#357EC7>buffer</font> to water.<br></p><p>Resuspend pellet by vortexing/by shaking vigorously.<br></li></p></ol><p><b>TOTAL TIME REQUIRED FOR THE COMPLETION OF THE PROTOCOL :<font color=#357EC7>~ 12 hrs, 25 mins</font></b></p></html> | ||
Latest revision as of 23:39, 19 November 2009
<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></ul><h2>Steps:</h2><ol><p><li>Measure out <b><font color=#357EC7>0.1</font></b> volume <font color=#357EC7>3M Sodium Acetate solution</font> into DNA sample.<br></li></p><p><li>Measure out <b><font color=#357EC7>1 µl</font></b> of <font color=#357EC7>Glycogen</font> into Eppendorf tube (1).<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><b>Option 1: </b>Store at <b><font color=#357EC7>-20°C</font></b> for <b><font color=#357EC7>12 hrs</font></b>(overnight).<br>(or)<br><b>Option 2: </b>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><b>Option 1: </b>Add <font color=#357EC7>water</font> to pellet.<br>(or)<br><b>Option 2: </b>Add <font color=#357EC7>buffer</font> to water.<br></p><p>Resuspend pellet by vortexing/by shaking vigorously.<br></li></p></ol><p><b>TOTAL TIME REQUIRED FOR THE COMPLETION OF THE PROTOCOL :<font color=#357EC7>~ 12 hrs, 25 mins</font></b></p></html>
