DNA Precipitation protocol

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(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><...)
Current revision (01:39, 20 November 2009) (view source)
 
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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</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>
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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</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>

Current revision

Solutions/reagents:

  • 3M Sodium Acetate solution
  • Glycogen
  • 95% EtOH
  • 70% EtOH
  • water
  • buffer
  • DNA sample

Equipment:

  • Centrifuge

Steps:

  1. Measure out 0.1 volume 3M Sodium Acetate solution into DNA sample.
  2. Measure out 1 µl of Glycogen into Eppendorf tube (1).
  3. Add 2 volumes 95% EtOH.
  4. Option 1: Store at -20°C for 12 hrs(overnight).
    (or)
    Option 2: Store at -80°C for 30 mins.

  5. Centrifuge at maximum speed for at least 15 mins at room temperature, gently aspirate out the supernatant and discard it.
  6. (Optional)
    Add 1 ml of 70% EtOH.
    Store at room temperature for 5 mins.

    (Optional)
    Centrifuge at maximum speed for 5 mins at room temperature, gently aspirate out the supernatant and discard it.

  7. Dry the pellet in air for 10 - 15 mins.
    Dry until all the liquid is gone.
  8. Option 1: Add water to pellet.
    (or)
    Option 2: Add buffer to water.

    Resuspend pellet by vortexing/by shaking vigorously.

TOTAL TIME REQUIRED FOR THE COMPLETION OF THE PROTOCOL :~ 12 hrs, 25 mins

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