Is the SMARTer Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian compatible with RNA purified from FFPE/PFA-fixed, or LCM cells?
Is the SMARTer Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian compatible with RNA purified from FFPE/PFA-fixed, or LCM cells?
The SMARTer Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian is compatible with purified RNA obtained from FFPE/PFA-fixed, and LCM cells, that does not contain any remnants of DNA, carrier or co-precipitant. RNA from FFPE or PFA-fixed cells must be decrosslinked and thoroughly treated with DNAse in order to remove all traces of DNA, since single-stranded DNA (often present in FFPE, PFA-fixed, and LCM cells) can be randomly primed and used as a template by the reverse transcriptase, thus incorporating DNA in to the final SMARTer Stranded v3 Pico library. If feasible, it is advisable to perform an additional DNAse I treatment of purified RNA eluate, followed by column purification, since DNA removal during bulk RNA purification (including on-column DNAse treatment) may not always be sufficient. Consider NucleoSpin RNA XS Total RNA Isolation User Manual (Section 5.4, ""rDNase digestion in the eluate""); NucleoSpin totalRNA FFPE - FFPE XS RNA Isolation User Manual (Section 5.3, ""DNA digestion in the RNA eluates""); NucleoSpin RNA Clean-up XS User Manual - 5.2 DNA digestion in crude RNA extracts and subsequent clean-up