IGEM:Harvard/2006/Salt-ethanol precipitation: Difference between revisions
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''Note: this protocol is preliminary'' | |||
==Small-scale separation using salt:ethanol precipitation== | ==Small-scale separation using salt:ethanol precipitation== | ||
*Add 0.25 mass equiv of 5 M NaCl (assume 1.19 g/mL density) | *Add 0.25 mass equiv of 5 M NaCl (assume 1.19 g/mL density) |
Latest revision as of 14:06, 20 July 2006
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.