Daniel:Protocols/ssDNA Column

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Revision as of 22:39, 31 July 2013 by >Djacobse (Created page with "=ssDNA Column Purification (Zymo)= '''Zymo: D7010 (20); D7011 (50)''' ==Protocol== 1. Add 2 volumes '''Binding Buffer''' to the sample; mix well 2. Transfer to '''IIC Col...")
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ssDNA Column Purification (Zymo)

Zymo: D7010 (20); D7011 (50)

Protocol

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