Gel purification

Gel purification kdorfman Tue, 03/29/2022 - 16:21

Zymoclean Gel DNA Recovery Kit Zymo Research D4002

  • Add 26 mL 95% EtOH to the 6 mL DNA Wash Buffer concentrate
  • Excise DNA fragment (x-tracta Gel Extraction Tool or razor blade)
  • Add 3X volume ADB (OR 300 uL ADB/mg gel)
  • Incubate 37-55C 5-10 minutes or until gel is dissolved
  • transfer melted agarose solution to Zymo Spin Column in a Collection Tube
  • centrifuge 10 - 16K Xg 30-60 sec
  • add 200 uL DNA Wash Buffer
  • Discard flow-through
  • Repeat with another 200 uL DNA Wash Buffer. Discard flow-through again
  • Add >= 6 uL DNA Elution Buffer (or water)
  • Put into clean 1.5 mL tube
  • Spin 30-60 sec to elute DNA.

Per rxn:

Reagent amount aliquot for 2 rxns for 3 rxns
ADB1 300 uL 640 uL 950 uL
DNA Wash Buffer 400 uL 840 uL 1250 uL
ENA Elution Buffer 6 uL 15 uL 22 uL (or water?)
  • Jewelry scale to weigh gel
  • Hot block 55 C for incubation and melting of agarose
  • gel excision tools
  • 2 mL tube for melting gel
  • vortex

  1. volume depends on size of excised gel piece ↩︎

Zymoclean Gel DNA Recovery Kit protocol