PSP Spin Stool DNA Kit - STRATEC Biomedical AG

TroubleshootingProblemComments and suggestionsClogged RTA Spin Filterinsufficient lysis and/ or too much starting materialincrease lysis timeincrease centrifugation speedreduce amount of starting materialLow amount or no DNA of extracted DNAsample stored incorrectlyinsufficient homogenization of stool sample inLysis Buffer P or in Stool DNA Stabilizerinsufficient lysisinsufficient mixing of the sample with BindingBuffer Ano alcohol added to the Wash Buffer I and IIsample should be stored at 4°C or – 20°Crepeat the DNA purification procedure with a new sample Besure to mix the sample and Lysis Buffer P or in Stool DNAStabilizer until the sample is thoroughly homogenizeduse Zirconia Beads II and vortex for homogenizationincrease lysis timereduce amount of starting materialoverloading of Spin Filter reduces yield!mix sample sufficient by pipetting up and down with BindingBuffer A prior to transfer the sample onto the RTA SpinFilter membranecheck that Wash Buffer I and Wash Buffer II concentrateswere diluted with correct volume of 96-100% ethanol.repeat the purification procedure with a new sampleDNA not eluted efficientlyto increase elution efficiency, pipet the preheated ElutionBuffer onto the center of the RTA Spin Filter and incubatethe column for 5 minutes at room temperature beforecentrifugationdo the elution steps twice.take higher volume of Elution Buffer.A260/A280 ratio for purified nucleic acids islowinefficient elimination of inhibitory substances dueto insufficient mixing with the InviAdsorb matrixinsufficient mixing with Lysis Buffer Pdecreased proteinase activityno Binding Buffer A added to the lysateWash Buffer I and Wash Buffer II preparedincorrectlyrepeat the DNA purification procedure with a new samplebe sure to mix the sample and InviAdsorb matrix until thesample is thoroughly homogenizedrepeat the procedure with a new samplebe sure to mix the sample and Lysis Buffer P immediatelyand thoroughly by pulse vortexingrepeat the DNA purification procedure with a new sampleand with Proteinase K for difficult cases use double volumeProteinase Krepeat the purification procedure with a samplecheck that Wash Buffer I and Wash Buffer II were dilutedwith 96–100% ethanoldo not use denatured alcohol, which contains othersubstances such as methanol or methylethylketonerepeat the purification procedure with a new samplePSP ® Spin Stool DNA Kit 0213PSP ® Spin Stool DNA Plus Kit 0213