Daniel:Protocols/ssDNA Column: Difference between revisions

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==Protocol==
==Protocol==


1. Add 2 volumes '''Binding Buffer''' to the sample; mix well
<ol>
2. Transfer to '''IIC Column''' and centrifuge at 14000 rpm for 1 minute; '''SAVE THE FLOW THROUGH'''
<li>Add 2 volumes '''Binding Buffer''' to the sample; mix well</li>
3. Add 1 volume 100% EtOH to flow through; mix well
<li>Transfer to '''IIC Column''' and centrifuge at 14000 rpm for 1 minute; '''SAVE THE FLOW THROUGH'''</li>
4. Transfer to '''IC Column''' and centrifuge at 14000 rpm for 1 minute; discard flow through
<li>Add 1 volume 100% EtOH to flow through; mix well</li>
5. Add 400 uL '''Prep Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through
<li>Transfer to '''IC Column''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li>
6. Add 700 uL '''Wash Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through
<li>Add 400 uL '''Prep Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li>
7. Add 400 uL '''Wash Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through
<li>Add 700 uL '''Wash Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li>
8. Centrifuge empty column at 14000 rpm for 2 minutes
<li>Add 400 uL '''Wash Buffer''' and centrifuge at 14000 rpm for 1 minute; discard flow through</li>
9. Transfer to empty 1.5mL centrifuge tube (low bind)
<li>Centrifuge empty column at 14000 rpm for 2 minutes</li>
10. Add ≥ 6 uL nfH2O and centrifuge at 14000 rpm for 1 minute
<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]

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Zymo: D7010 (20); D7011 (50)

Protocol[edit]

  1. Add 2 volumes Binding Buffer to the sample; mix well
  2. Transfer to IIC Column and centrifuge at 14000 rpm for 1 minute; SAVE THE FLOW THROUGH
  3. Add 1 volume 100% EtOH to flow through; mix well
  4. Transfer to IC Column and centrifuge at 14000 rpm for 1 minute; discard flow through
  5. Add 400 uL Prep Buffer and centrifuge at 14000 rpm for 1 minute; discard flow through
  6. Add 700 uL Wash Buffer and centrifuge at 14000 rpm for 1 minute; discard flow through
  7. Add 400 uL Wash Buffer and centrifuge at 14000 rpm for 1 minute; discard flow through
  8. Centrifuge empty column at 14000 rpm for 2 minutes
  9. Transfer to empty 1.5mL centrifuge tube (low bind)
  10. Add ≥ 6 uL nfH2O and centrifuge at 14000 rpm for 1 minute