Team:UNIPV-Pavia/Protocols/Precipitation

From 2008.igem.org

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*'''absolute ethanol'''
*'''absolute ethanol'''
*'''ethanol 70%'''
*'''ethanol 70%'''
 +
*'''ddH2O'''
*'''previously cut plasmid'''
*'''previously cut plasmid'''
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*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
-
*decant supernatant
+
*Decant supernatant
-
*Add 50 l of 70% ethanol
+
*Add 50 µl of 70% ethanol
*Centrifuge at 13000 rpm, 4°C for 20 min
*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 µl ddH2O
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<br>

Latest revision as of 13:30, 1 July 2008

Home.jpg Home Unipv logo.jpg The Team And.jpg The Project Safety.jpg Biological Safety Dna.png Parts Submitted to the Registry
Laptop.jpg Dry Lab Pipette.jpg Wet Lab Math.gif Modeling Note.jpg Protocols Notebook.gif Activity Notebook


The protocols we used


DNA precipitation with sodium acetate

(estimated time: 1 hour)

Materials needed:

  • sodium acetate
  • absolute ethanol
  • ethanol 70%
  • ddH2O
  • previously cut plasmid


  • Add 1/10 digestion volume of sodium acetate 3 M
  • Add 2.5 digestion 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 µl ddH2O