Difference between revisions of "Template:SBB-Protocols Zymo2"

From OpenWetWare
Jump to: navigation, search
 
Line 6: Line 6:
 
#Add 500uL of Ethanol and pipette up and down to mix
 
#Add 500uL of Ethanol and pipette up and down to mix
 
#spin through (15s), discard waste.
 
#spin through (15s), 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 (15s), discard waste.
 
#spin through (15s), 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 (spin 60s) into a fresh Eppendorf tube
 
#elute with water (spin 60s) into a fresh Eppendorf tube

Latest revision as of 09:10, 2 February 2012

Small-Frag Zymo Cleanup

The following procedure removes the polymerase, dNTPs, buffer, and most of the oligonucleotides from a PCR reaction for fragments smaller than 300bp. It also will remove the buffer and restriction enzymes from a restriction digest reaction.

  1. Add 100 uL of Zymo ADB buffer (brown bottle) to the reaction.
  2. Transfer into the Zymo column (small clear guys)
  3. Add 500uL of Ethanol and pipette up and down to mix
  4. spin through (15s), discard waste.
  5. Add 200 uL of Zymo Wash Buffer (which is basically 70% ethanol)
  6. spin through (15s), discard waste.
  7. Add 200 uL of Zymo Wash Buffer
  8. spin through, discard waste.
  9. spin for 90 seconds, full speed to dry.
  10. elute with water (spin 60s) into a fresh Eppendorf tube