Cat # SL100388 Store at 4~23 0 C

1. DiatoCLEAN™ DNA Purification Kit ------Quick Protocol Small 300 Preps Large 600 Preps 15875 Gaither Drive Gaithersburg, MD 20877 FAX. 301-560-4919 TEL. 301-330-5966 Toll-Free. 1-866-918-6812 Email: info@signagenlabs.com Web: www.signagenlabs.com Cat # SL100388 Store at 4~23 0C This product is for laboratory research ONLY and not for diagnostic use 6. Resuspend the DiatoCLEAN™ powder/DNA pellet with TE buffer or water by vortexing. Use at least twice the volume of the original DiatoCLEAN ™ Slurry used. Incubate the tube at 75 0C for 5 minutes. Vortex and centrifuge at 13,000 rpm for 30 seconds and carefully remove the supernatant containing the eluted DNA and place in a new tube. Higher the elution efficiency is achieved using a large volume and performing 2-3 elution cycles with the new portion of eluant. Eluted DNA can be used immediately in enzymatic or other manipulations. Note: Incubation of DiatoCLEAN™ powder/DNA pellet with TE buffer water at 75 0C for 5 minutes is essential for good DNA recovery. II. Recovering DNA from Polymerase Chain Reactions 1. Remove the mineral oil (if any) from PCR tube. Measure the volume of PCR product.Add at least 3 volumes of DNA Binding Buffer and mix thoroughly.2. Add the resuspended DiatoCLEAN™ Slurry (for 1.0 µg DNA add 3.0 µl slurry). Incubate for 5 minutes at 75 0C by mixing occasionally to keep DiatoCLEAN™ particle resuspended in the solution. 4. Spin at 13,000 rpm for 30 seconds to form a pellet. Remove the supernatant. 5. Add 500 ml of ice cold DNA Wash Buffer to the pellet. Resuspend the pellet in the wash buffer by gentle pipetting. Centrifuge at 13,000 rpm for 30 seconds and discard the supernatant. Repeat the wash procedure once again. During each wash the pellet should be resuspended completely. Note: Ice-cold DNA Wash Buffer is essential for good DNA recovery6. After the supernatant from the last wash has been removed, spin the tube again and remove the remaining liquid with pipette tip. Air dry the DiatoCLEAN™ powder/DNA pellet for 5 minutes. Note: Try to avoid overdrying DNA, which may cause poor DNA recovery 7. Resuspend the DiatoCELAN™ powder/DNA with TE buffer or water by vortexing. Use at least twice the volume of the original DiatoCLEAN™ Slurry used.Incubate the tube at 75 0C for 5 minutes. Vortex and centrifuge at 13,000 rpm for 30 seconds and carefully remove the supernatant containing the eluted DNA and place in a new tube. Higher elution efficiency can be achieved using a large volume and performing 2-3 elution cycles with the new portion of eluant. Eluted DNA can be used immediately in enzymatic or other manipulations. Storage: Upon arrival store this product at 4 0C ~room temperature. For long term, store the product at 4 0C. Product shipped at ambient temperature. Description: DiatoCLEAN™ DNA Purification Kit is formulated to use super fine diatomaceous earth to specifically bind and purify DNA. As diatomaceous earth particle is very fine and usually has very irregular surface, leading to much higher binding capacity than spherical silica particle. The purified DNA is OK for digestion, ligation, transformation, sequencing, etc. Advantages of DiatoCLEAN™ DNA Purification Kit: • DNA specific--------diatomaceous earth exclusively binds DNA (not RNA or protein) with wide size range (> 100 bp)• No addition of sodium iodide which undergoes decomposition in aqueous solution and may modify DNA molecules during purification • Fast purification------DNA isolation and purification take less than 20 minutes • TAE and TBE gels work equally good • High efficiency with more than 90 % DNA recovery Applications:• Purify DNA from any type of agarose gels • Concentrate DNA for changing buffer, desalting, etc• Remove proteins after restriction enzyme treatment, dephosporylation with BAP or CIAP etc• Remove unincorporated nucleotides, primer, primer-dimers and enzymes from a labeling reaction or PCR• Remove an excess of linkers after ligation• Remove residual phenol, chloroform, or ethidium bromide.• Purify plasmid DNA free of RNA from bacterial lysates Instructions:I. Recovering DNA from Agarose Gel1. Run the agarose gel to separate DNA fragments. Cut agarose gel band containing the desired DNA. Determine an approximate volume of gel slice by weight (1 mg equals approximately 1 µl) and place the slice into a plastic tube. Note: Minimize the time of UV exposure as much as it is possible. 2. Add at least 3 volumes of DNA Binding Buffer. Incubate 5 minutes at 75 0C to dissolve agarose by mixing occasionally to promote agarose solubilization. If the solubilization is incomplete, incubate another 5 minutes. 3. Add appropriate volume of the resuspended DiatoCLEAN™ Slurry (1.0 µg DNA add 3.0 µl slurry). Incubate for 5 minutes at 75 0C. Mix occasionally to keep the slurry resuspended in the solution to form slurry-DNA complex, do not vortex. 4. Spin DiatoCLEAN™ slurry/DNA complex at 13,000 rpm for 30 seconds to form a pellet . Remove the supernatant. Add 500 µl of DNA Binding Buffer to the pellet and resuspend it by gentle pipetting. Spin again at 13,000 rpm for 30 seconds to form pellet. Remove the supernatant. 5. Add 500 µl of ice cold DNA Wash Buffer to the pellet. Resuspend the pellet in the DNA Wash Buffer by gentle pipetting (do not vortex high molecular weight DNA). Centrifuge at 13,000 rpm for 30 seconds and discard the supernatant. Repeat the wash procedure once again. Briefly spin the tube again and remove the remaining liquid with pipette tip. Air dry the pellet for 5 minutes.Note: Ice-cold DNA Wash Buffer is essential for good DNA recovery. Try to avoid overdrying DNA, which may cause poor DNA recovery 2006 SignaGen Laboratories