Daniel:Protocols/ssDNA Column: Difference between revisions
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==Protocol== | ==Protocol== | ||
<ol> | |||
<li>Add 2 volumes '''Binding Buffer''' to the sample; mix well</li> | |||
<li>Transfer to '''IIC Column''' and centrifuge at 14000 rpm for 1 minute; '''SAVE THE FLOW THROUGH'''</li> | |||
<li>Add 1 volume 100% EtOH to flow through; mix well</li> | |||
<li>Transfer to '''IC Column''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li> | |||
<li>Add 400 uL '''Prep Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li> | |||
<li>Add 700 uL '''Wash Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li> | |||
<li>Add 400 uL '''Wash Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li> | |||
<li>Centrifuge empty column at 14000 rpm for 2 minutes</li> | |||
<li>Transfer to empty 1.5mL centrifuge tube (low bind)</li> | |||
<li>Add ≥ 6 uL nfH2O and centrifuge at 14000 rpm for 1 minute</li> | |||
</ol> |
Latest revision as of 21:05, 16 August 2016
ssDNA Column Purification (Zymo)[edit]
Zymo: D7010 (20); D7011 (50)
Protocol[edit]
- Add 2 volumes Binding Buffer to the sample; mix well
- Transfer to IIC Column and centrifuge at 14000 rpm for 1 minute; SAVE THE FLOW THROUGH
- Add 1 volume 100% EtOH to flow through; mix well
- Transfer to IC Column and centrifuge at 14000 rpm for 1 minute; discard flow through
- Add 400 uL Prep Buffer and centrifuge at 14000 rpm for 1 minute; discard flow through
- Add 700 uL Wash Buffer and centrifuge at 14000 rpm for 1 minute; discard flow through
- Add 400 uL Wash Buffer and centrifuge at 14000 rpm for 1 minute; discard flow through
- Centrifuge empty column at 14000 rpm for 2 minutes
- Transfer to empty 1.5mL centrifuge tube (low bind)
- Add ≥ 6 uL nfH2O and centrifuge at 14000 rpm for 1 minute