Gel purification
Gel purification kdorfman Tue, 03/29/2022 - 16:21Zymoclean 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
-
volume depends on size of excised gel piece ↩︎