Daniel:Protocols/ssDNA Column: Difference between revisions

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=ssDNA Column Purification (Zymo)=
=ssDNA Column Purification (Zymo)=
[[Daniel Jacobsen|Back to Main]]


'''Zymo: D7010 (20); D7011 (50)'''
'''Zymo: D7010 (20); D7011 (50)'''

Revision as of 22:40, 31 July 2013

ssDNA Column Purification (Zymo)

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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