Brandon:Protocols/AMPure beads purification: Difference between revisions
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>Bsos (Created page with '==AMPure beads purification protocol== *Purification with AMPure beads *1. 50ul beads to 50ul reaction (or 1 volume to 1 volume), mix 10 times, R.T for 8min *2. Place on magn…') |
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*1. 50ul beads to 50ul reaction (or 1 volume to 1 volume), mix 10 times, R.T for 8min | *1. Add 50ul beads to 50ul reaction (or 1 volume to 1 volume), mix 10 times, R.T for 8min | ||
*2. Place on magnetic stand. | *2. Place on magnetic stand. | ||
*3. After 5 minutes, remove supernatant, wash with 180ul fresh made 80% EtOH twice. | *3. After 5 minutes, remove supernatant, wash with 180ul fresh made 80% EtOH twice. | ||
*4. Remove from magnetic stand. Air-dry for 15min, re-suspend with 25ul Qiagen EB (or nuclease free H2O), mix 10 times, let sit at RT for 2min, then place in magnetic stand. | *4. Remove from magnetic stand. Air-dry for 15min, re-suspend with 25ul Qiagen EB (or nuclease free H2O), mix 10 times, let sit at RT for 2min, then place in magnetic stand. | ||
*5. After 2 minutes, transfer 20ul DNA from beads (leave a little so don't pick up beads), 1ul left in tube for Qubit measurement | *5. After 2 minutes, transfer 20ul DNA from beads (leave a little so don't pick up beads), 1ul left in tube for Qubit measurement |
Revision as of 21:34, 6 April 2012
AMPure beads purification protocol
- Purification with AMPure beads
- 1. Add 50ul beads to 50ul reaction (or 1 volume to 1 volume), mix 10 times, R.T for 8min
- 2. Place on magnetic stand.
- 3. After 5 minutes, remove supernatant, wash with 180ul fresh made 80% EtOH twice.
- 4. Remove from magnetic stand. Air-dry for 15min, re-suspend with 25ul Qiagen EB (or nuclease free H2O), mix 10 times, let sit at RT for 2min, then place in magnetic stand.
- 5. After 2 minutes, transfer 20ul DNA from beads (leave a little so don't pick up beads), 1ul left in tube for Qubit measurement