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Problem/ Probable cause Comments and suggestions DNA-contamination too much starting material reduce amount of starting material DNase digestion of the eluate containing the total RNA no optimal homogenization of DNA binding carrier shake Lysis Solution R carefully before use RNA degraded inappropriate handling of the starting material the RNA purification protocol should be performed quickly (see also “General notes on handling RNA”, page 34) cell pellets stored at - 80°C for later processing should be immediately frozen after cell harvest by liquid nitrogen treatment or under Lysis Solution Lysis Solution does not contain DTT ensure that DTT has been added to the Lysis Solution R RNase contamination check for RNase contamination of buffers, be certain not to introduce any RNase throughout the procedure or during further handling for analysis see appendix , ”General Notes on handling RNA” (page 34) total RNA does not perform well in downstreamapplications (RT-PCR) ethanol carryover during elution salt carryover during elution increase g-force or centrifugation time when drying the RTA Spin Filter ensure that Wash Buffer R1 and R2 are at room temperature check up Wash Buffer R1 and R2 for salt precipitates, if there are any precipitates solve these precipitates by careful warming 26 InviTrap® Spin Cell RNA Mini Kit 0515
