IGEM:Harvard/2006/Salt-ethanol precipitation

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Note: this protocol is preliminary

Small-scale separation using salt:ethanol precipitation

  • Add 0.25 mass equiv of 5 M NaCl (assume 1.19 g/mL density)
    • example: 100 μL * 0.25 / 1.19 = 21 μL
  • Add 0.5 mass equiv of ethanol (assume density of 0.79 g/mL)
    • example: 100 μL * 0.5 / 0.79 = 63.3 μL
  • Incubate on ice 10 min
  • Centrifuge at 15000 rpm for 10 min at 4°C
  • Carefully remove supernatant with pipette
  • Resuspend in 50 μL 1x folding buffer

Stock is now concentrated 2x, but we probably suffered some losses. Run a gel and check spot-density of the bands to determine yield.

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