What RNA purification kits are recommended for compatibility with the SMART-Seq Stranded cDNA synthesis?
What RNA purification kits are recommended for compatibility with the SMART-Seq Stranded cDNA synthesis?
We recommend the NucleoSpin RNA XS kit offering on-column DNAse I treatment as well as an optional DNAse I treatment of RNA eluate, followed by column purification. To ensure a complete removal of DNA, essential for the highly sensitive SMART-Seq Stranded cDNA synthesis, it is advisable to perform an additional DNAse I treatment of purified RNA eluate, since single-stranded DNA (present in the RNA prep or generated during the fragmentation step) can be randomly primed and used as a template by the reverse transcriptase, thus incorporating genomic DNA sequences into the final SMART-Seq Stranded library. Do not use RNA purification kits omitting DNAse treatment. Do not use any carrier or co-precipitant during RNA purification, since it may distort RNA concentration and interfere with cDNA synthesis.