Daniel:Protocols/ssDNA Column

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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