The Ultra-Sep^® Gel Extraction kit is the most economical and convenient kit available for the isolation of DNA fragments from agarose gel.

Principle

By combining the silica particle method and Omega Bio-Tek's unique buffer system, this kit offers an easy and flexible protocol. The DNA band of interest is excised from the gel, dissolved in Ultra-Sep^® Binding Buffer, and then mixed with Ultra-Sep^® Beads that bind DNA. Following a rapid wash step, DNA is eluted with de-ionized water (or low salt buffer) and is ready for other applications. The product is suitable for ligations, PCR amplification, restriction digestion, and various labeling reactions.

Procedure

Perform agarose gel/ethidium bromide electrophoresis to fractionate DNA fragments. Carefully excise the fragment of interest while visualizing under UV light. Assuming a density of 1 g ml-1 of gel, add 1ml of Binding Buffer per gram of gel and incubate to dissolve agarose. Apply the DNA/agarose solution to 10 µl Ultra-Sep™ beads and incubate at room temperature for 10 minutes. Centrifuge to pellet the beads. Discard the liquid and wash the pelleted beads once with Ultra-Sep™ binding buffer and then twice with wash buffer diluted with ethanol to remove impurities. Completely remove the liquid from micro-centrifuge tube and air-dry the pellet for 10-15 minutes. Re-suspend the pellet in sterile de-ionized water or TE buffer and incubate at 50-55oC in a water bath for 5 minutes. Centrifuge to pellet the Ultra-Sep™ beads and transfer the supernatant containing pure DNA to a clean tube.

Downstream Applications

DNA fragments purified with the Ultra-Sep™. DNA Gel Extraction Kit are ready for direct use in all applications, including :

• Ligations
• Restriction digestion
• Labeling
• PCR Sequencing

DNA fragments recovery from agarose using Ultra-Sep™ Gel Extraction Kit.
DNA fragments are : 50bp (lane 1), 100bp (lane 2), 200bp (lane 3),
300bp (lane 4), 400bp (lane 5). M: Low molecular weight marker.