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=== 4. ZymoBead™ Genomic DNA kit === : -This protocol is for whole blood, serum and plasma samples ==== Procedures ==== : -Before starting, add beta-mercaptoethanol to Genomic Lysis Buffer to a final dilution of 0.5% (v/v). Vortex ZymoBead to resuspend : <u>Preparing lysate and binding DNA</u> :: - Add 200ul of '''Genomic Lysis Buffer''' to '''50ul samples''' in 1.5mL tube :: - Add 10ul '''ZymoBeads'''. Mix by inversion :: - Incubate at RT for 5 min :: - Centrifuge at 1,500xg for 1 min :: - Carefully remove the spnt without disturbing the bead pellet :: - Add 200ul of '''Genomic Lysis Buffer to the ZymoBeads :: Resuspend the beads by pipetting up & down :: Centrifuge at 1,500xg for 1 min :: - Carefully remove the spnt without disturbing the bead pellet : <u>Washing DNA</u> :: - Add '''200ul''' of '''DNA Pre-Wash Buffer''' to the ZymoBeads. :: - Resuspend the pellet, transfer to a new tube, centrifuge at 1,500xg for 1min :: - Carefully remove the spnt without disturbing the bead pellet :: Add '''500ul''' of '''g-DNA Wash Buffer''' to the ZymoBeads. :: - Resuspend the pellet and centrifuge at 1,500xg for 1 min. :: - Carefully remove the spnt without disturbing the bead pellet :: Re-centrifuge briefly and remove the residual wash buffer : <u>Eluting DNA</u> :: - Add '''>35ul''' of '''Elution Buffer''' or H2O and resuspend the pellet by pipetting up & down. :: - Centrifuge at 10,000xg for 1 min :: - Collect the spnt to the microcentrifuge tube.
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