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Revision as of 21:11, 27 September 2011

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TITLE=Plasmid Extraction| BODY=

Gel extraction

This protocol is utilized in accordance to the manufacturer's protocol from Omega E.Z.N.A (EaZy Nucleic Acid Isolation)

1. Place gel on the UV box
2. Carefully extract the fragment suspended in the gel>/li>
3. Mass gel fragments
4. Place fragment into a 1.5 mL tube and add 4 µL of H2O
5. Volume of water added to volume of gel is 200% however if fragment it small 1 mL of water will suffice
6. Remove H2O
7. Add equal amounts of H2O and Binding Buffer (XP2) to the gel
8. Incubate mixture at 55 degrees for 7 mins
9. Mix with vortex for 2 mins
10. Place in the HiBind DNA Mini Column in the 2 mL tube
11. Add 700 µL at 10,000xg for 1 min
12. Discard liquid
13. Add 300 µL Binding Buffer (XP2) into the HiBind DNA Mini Column and spin down at 10,000xg for 1 min
14. Discard liquid
15. Wash the column with 700 µL of SPW buffer with added ethanol and spin down at 10,000xg for 1 min
16. Discard liquid
17. Wash the column with 700 µL of SPW buffer again and spin down at 10,000xg for 1 min
18. Discard the liquid
19. Spin down the column at 13,000xg for 1 min to dry the column
20. Elute in 50 µL of H2O and wait 1 min
21. Spin down the column at 13,000xg for 1 min to dry the column
22. Use a spectrophotometer to measure the concentration and the purity of your plasmid