Maheshri:EtOHppt: Difference between revisions

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==Protocol for Nucleic Acid Precipitation from Diluted Solutions==
<Font size="+1">Protocol for Nucleic Acid Precipitation from Diluted Solutions</font>


1. Add 1/10 volume of 3M sodium acetate (or 2M sodium chloride, or 5M ammonium acetate) to DNA solution.
#Add 1/10 volume of 3M sodium acetate (or 2M sodium chloride, or 5M ammonium acetate) to DNA solution.
 
#Add Glycogen to a final 0.05-1µg/µl concentration. Use up to 1µl of Glycogen per 20µl of the solution.
2. Add Glycogen to a final 0.05-1µg/µl concentration. Use up to 1µl of Glycogen per 20µl of the solution.
#Add 1 volume of isopropanol (or 2.5 volumes of ethanol) to the solution. Mix gently but thoroughly. Ethanol is recommended for precipitation of smaller than 200 b/bp RNA/DNA fragments.
 
#Incubate the mixture at -20°C for up to 60min, or at -70°C for 30min.
3. Add 1 volume of isopropanol (or 2.5 volumes of ethanol) to the solution. Mix gently but thoroughly. Ethanol is recommended for precipitation of smaller than 200 b/bp RNA/DNA fragments.
#Centrifuge the mixture for 10-15min at 10,000rpm. Discard the supernatant.
 
#Rinse the pellet with cold 70% ethanol. Air-dry the pellet.
4. Incubate the mixture at -20°C for up to 60min, or at -70°C for 30min.
#Dissolve DNA in Water, nuclease-free <http://www.fermentas.com/catalog/reagents/water.htm> (#R0581) or TE buffer.
 
#Dissolve RNA in DEPC-treated Water <http://www.fermentas.com/catalog/reagents/depc.htm> (#R0601).
5. Centrifuge the mixture for 10-15min at 10,000rpm. Discard the supernatant.
 
 
''Using only sodium acetate and ethanol works well - CJZ 6/24/10''
6. Rinse the pellet with cold 70% ethanol. Air-dry the pellet.
 
7. Dissolve DNA in Water, nuclease-free <http://www.fermentas.com/catalog/reagents/water.htm> (#R0581) or TE buffer.
 
8. Dissolve RNA in DEPC-treated Water <http://www.fermentas.com/catalog/reagents/depc.htm> (#R0601).

Latest revision as of 12:35, 24 June 2010

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Protocol for Nucleic Acid Precipitation from Diluted Solutions

  1. Add 1/10 volume of 3M sodium acetate (or 2M sodium chloride, or 5M ammonium acetate) to DNA solution.
  2. Add Glycogen to a final 0.05-1µg/µl concentration. Use up to 1µl of Glycogen per 20µl of the solution.
  3. Add 1 volume of isopropanol (or 2.5 volumes of ethanol) to the solution. Mix gently but thoroughly. Ethanol is recommended for precipitation of smaller than 200 b/bp RNA/DNA fragments.
  4. Incubate the mixture at -20°C for up to 60min, or at -70°C for 30min.
  5. Centrifuge the mixture for 10-15min at 10,000rpm. Discard the supernatant.
  6. Rinse the pellet with cold 70% ethanol. Air-dry the pellet.
  7. Dissolve DNA in Water, nuclease-free <http://www.fermentas.com/catalog/reagents/water.htm> (#R0581) or TE buffer.
  8. Dissolve RNA in DEPC-treated Water <http://www.fermentas.com/catalog/reagents/depc.htm> (#R0601).

Using only sodium acetate and ethanol works well - CJZ 6/24/10

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