Template:SBB-Protocols Zymo3

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(Zymo Gel Purification)
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#transfer into the Zymo column inside a collection tube (small clear guys)
#transfer into the Zymo column inside a collection tube (small clear guys)
#spin through, discard waste.
#spin through, discard waste.
-
#Add 200 uL of PE buffer (which is basically 70% ethanol)
+
#Add 200 uL of A4 Wash Bbuffer (which is basically 70% ethanol)
#spin through, discard waste.
#spin through, discard waste.
-
#Add 200 uL of PE buffer
+
#Add 200 uL of A4 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 8.5 uL of water into a fresh Eppendorf tube
+
#elute with water into a fresh Eppendorf tube

Revision as of 23:14, 21 February 2011

Zymo Gel Purification

  • All spins until the drying step are 15 second full speed spins.
  1. cut out bands minimizing extra gel matter.
  2. put in ependorf tube and add 600uL of Zymo ADB buffer (brown bottle).
  3. heat at 55, shake and/or vortex until the gel has dissolved.
  4. If the DNA is <300bp add 250uL of isopropanol
  5. transfer into the Zymo column inside a collection tube (small clear guys)
  6. spin through, discard waste.
  7. Add 200 uL of A4 Wash Bbuffer (which is basically 70% ethanol)
  8. spin through, discard waste.
  9. Add 200 uL of A4 Wash Buffer
  10. spin through, discard waste.
  11. spin for 90 seconds, full speed to dry.
  12. elute with water into a fresh Eppendorf tube
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