1.Perform agarose gel/ethidium bromide electrophoresis to fractionate DNA fragments. Any type or grade of agarose may be used. However, it is strongly recommended that fresh TAE buffer or TBE buffer be used as running buffer. Do not reuse running buffer as its pH will increase and reduce yields.

2. When adequate separation of bands has occurred, carefully excise the DNA fragment of interest using a wide, clean, sharp scalpel. Minimize the size of the gel slice by removing extra agarose.

3. Determine the appropriate volume of the gel slice by weighing it in a clean 2mL

microcentrifuge tube. Add 1 volume Binding Buffer(XP2). Incubate at 55-65℃ for 7 minutes or until the gel has completely melted Vortex or shake the tube every 2-3 minutes.

4. Insert a HiBind DNA Mini Column in a 2 ml Collection Tube.

5. Add no more than 700μL DNA/agarose solution from Step 5 to the HiBind DNA Mini Column.Centrifuge at 13,000 x g for 1 minute at room temperature

6. Discard the filtrate and reuse collection tube.

7. Repeat Steps 7-9 until all of the sample has been transferred to the column.

8. Add 300 μL Binding Buffer (XP2). Centrifuge at maximum speed(13,000 x g) for 1 minute at room temperature. Discard the filtrate and reuse collection tube.

Note: SPW Wash Buffer must be diluted with 100% ethanol prior to use.

9. Centrifuge at maximum speed for 1 minute at room temperature. Discard the filtrate and reuse collection tube.

10. Centrifuge the empty HiBind DNA Mini Column for 2 minutes at maximum speed to

dry the column matrix

Note: It is important to dry the HiBind DNA Mini Column matrix before elution.

Residual ethanol may interfere with downstream applications.

11. Transfer the HiBind DNA Mini Column to a clean 1.5 mL microcentrifuge tube.Add 15-30 μL Elution Buffer or deionized water directly to the center of the column membrane.

12. Let sit at room temperature for 2 minutes. Centrifuge at maximum speed for 1 minute.

13. Store DNA at-20°C.