Do you have any recommendations for RNA extractions? What about for serum and plasma samples?

We highly recommend using Lexogen's SPLIT RNA Extraction Kit (Cat. No. 008), which recovers the complete RNA size range, including small RNAs (<200 nt and down to 17 nt). Additionally, large and small RNA-enriched fractions can be extracted by following a supplemental protocol.

When isolating total RNA for input, ensure the protocol can recover small RNAs (including miRNAs). Many commercial products are using silica-based purification technology that may exclude small RNAs. Please consult the manufacturer's specifications.

Alternatively, the TraPR Small RNA Isolation Kit (Cat. No. 128) offers a unique technology for enriching functional, physiologically relevant sRNAs that are associated with RISC complexes, even in uncharacterized samples, and thus simplifies downstream analysis and reduces sequencing costs. Please refer to the TraPR Small RNA Isolation Kit User Guide to find the recommended input amounts for your sample type.

RNA extraction from challenging samples:

• For FFPE samples, we recommend using Lexogen´s SPLIT One-Step FFPE RNA Extraction Kit (Cat. No. 236) which preserves small RNAs for downstream sRNA-Seq applications. To increase the RNA concentration in the eluate, precipitation or column-based clean-up can be performed after the extraction.

• For exosome samples derived from serum and plasma, we recommend isolating RNA with Qiagen exoRNeasy kits. Use 5 µl of total RNA eluate as the starting material, provided that 1 ml of plasma or serum was used for the exosomal RNA extraction.