Template:SBB-Protocols Zymo1

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Current revision (12:09, 2 February 2012) (view source)
 
Line 6: Line 6:
#Transfer into the Zymo column (small clear guys)
#Transfer into the Zymo column (small clear guys)
#spin through, discard waste.
#spin through, discard waste.
-
#Add 200 uL of A4 Wash Buffer (which is basically 70% ethanol)
+
#Add 200 uL of Zymo Wash Buffer (which is basically 70% ethanol)
#spin through, discard waste.
#spin through, discard waste.
-
#Add 200 uL of A4 Wash Buffer
+
#Add 200 uL of Zymo Wash Buffer
#spin through, discard waste.
#spin through, discard waste.
#spin for 90 seconds, full speed to dry.
#spin for 90 seconds, full speed to dry.
#elute with water into a fresh Eppendorf tube, use the same volume of water as the volume of the original reaction
#elute with water into a fresh Eppendorf tube, use the same volume of water as the volume of the original reaction

Current revision

Regular Zymo Cleanup

The following procedure removes the polymerase, dNTPs, buffer, and most of the oligonucleotides from a PCR reaction. It also will remove the buffer and restriction enzymes from a restriction digest reaction.


  1. Add 180 uL of Zymo ADB buffer (brown bottle) to a 33uL or 50uL reaction.
  2. Transfer into the Zymo column (small clear guys)
  3. spin through, discard waste.
  4. Add 200 uL of Zymo Wash Buffer (which is basically 70% ethanol)
  5. spin through, discard waste.
  6. Add 200 uL of Zymo Wash Buffer
  7. spin through, discard waste.
  8. spin for 90 seconds, full speed to dry.
  9. elute with water into a fresh Eppendorf tube, use the same volume of water as the volume of the original reaction
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