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E-Z 96® MagSi DNA Tissue DNA Kit for Buccal Swabs The following protocol is designed for isolating DNA from buccal swabs with magnetic beads. Kit Contents Product Number M6229-00 M6229-01 M6229-02 Purification Times 1 x 96 Preps 4 x 96 Preps 20 x 96 Preps 500 :l Micro Plate 1 4 20 1.2ml Process Plate 1 4 20 1. Place the swab into each well of 96 well plate(no provided). 2. Prepare a Proteinase K/Buffer TL working solution by mixing 20 :l Proteinase K with 250:l Buffer TL for each sample. For each 96 sets of sample, prepare the Protease/TL stock working solution by mix 2.0mL protease with 25mL Buffer TL. Pipet 250 :l protease/TL working solution into each well. Seal the plate properly. 8-strip caps 10 x 8 35 x 8 170 x 8 MagSi DNA Particles 2.2 ml 9 ml 45ml Buffer TL 20 ml 90 ml 2 x 180 ml Buffer MSL 20 ml 90 ml 2 x 180 ml Incubate at 56°C for 1 hours. Mix occasionally during the incubation by rotating the plate gently. Make sure the samples are completely lysed. Buffer MP 30 ml 100 ml 8 x 100 ml 2 ml 8 ml 32 ml 3. Proteinase K 4. Spin at 3,000-5,000 x g for 1 minutes at room temperature. RNase A 550 :l 2.1 ml 10 ml 5. Transfer 150 :l of the Sample into each well of Plate (provided). Elution Buffer 100 ml 2 x 200 ml 8 x 250 ml Add 150:l Buffer MSL, 20 :l MagSi DNA Paritcles and 220 :l absolute ethanol to the lysate. Mix the sample by pipetting up and down 20-30 times to mix well or shaking to mix well. Note: Buffer MSL, MagSi DNA Particles and absolute ethanol can be premixed. The mixture can be stored at room temperature for 3 hours. 1 1 1 6. 7. Place the Plate on a magnetic separation device (MSD-01) to magnetize the MagSi DNA particles for 3 min. The liquid should be cleared after all the magnetic beads are pelleted on the corner of the each well adjecent to the magnet. 8. Aspirate and discard the cleared supernatant. 9. Remove the Plate containing the MagSi DNA particles from the magnetic separation device. Add 500:l Buffer MP diluted with ethanol into each well. Note: Buffer MP must be diluted with absolute ethanol (96-100%) before use in this protocol. Instruction Booklet ® 10. Resuspend MagSi DNA particles pellet by pipetting up and down 20 times or shaking to mix well. 10 3