Gel Extraction Procedure

Excising and Dissolving the Gel

1. Excise a minimal area of gel (up to 400 mg) containing the DNA fragment.

2. Add Gel Solubilization Buffer (L1) to the excised gel in the tube size as indicated:

3. Place the tube with the gel slice and Buffer L1 into a 50oC water bath or heat block. Incubate the tube at 50oC for 15 minutes. Invert the tube every 3 minutes to mix.

4. Purify DNA using Centrifugation as described below.

Purifying DNA using Centrifugation

1. Load. Place a column into a 2-mL Receiver Tube. Pipet the dissolved gel piece onto the column. Centrifuge the column at >12,000 × g for 1 minute. Discard the flow-through and place the column into the 2-mL Receiver Tube.

2. Wash. Add 500 μL Wash Buffer (L2) containing ethanol to the column. Centrifuge the column at >12,000 × g for 1 minute. Discard the flow-through and place the column into the Receiver Tube. Centrifuge the column at maximum speed for 1 minute.

3. Elute. Place the column into a clean 1.5 mL microcentrifuge tube. Add 50 μL of TE Buffer to the column. Incubate the tube for 1 minute at room temperature. Centrifuge the tube at >12,000 × g for 2 minutes.

4. Store. The elution tube contains the purified DNA. Store the purified DNA at −4°C for immediate use or at−20°C for long-term storage.