IGEM:Harvard/2006/Salt-ethanol precipitation

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.