Gel Extraction

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

Reagents and Materials

• Omega E.Z.N.A (EaZy Nucleic Acid Isolation) kit
• Agarose gel
• Blade

Protocol

1. Place gel on the UV box
2. Carefully extract the fragment suspended in the gel
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°C 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