NEXTprep™ Small RNA Isolation Kit by Bioo Scientific

The NEXTprep™ Small RNA Isolation Kit is designed for enrichment of small RNA from eukaryotic cells and tissues. The small RNA recovered has been verified as suitable for making small RNA libraries for next generation sequencing (NGS) using the NEXTflex Small RNA Sequencing Kit. The NEXTprep Small RNA Isolation Kit includes BiooPure™ RNA Extraction Reagent for initial disruption of tissues or cells. BiooPure is a single-phase reagent that contains guanidinium thiocyanate, a powerful chaotropic agent effective for rapidly inactivating nucleases, and phenol, an organic solvent used to denature and separate proteins and DNA from RNA. The kit also includes silica filter spin basket assemblies and tubes for final purification of small RNA by solid phase extraction. The kit allows recovery of small RNAs approximately 100 bases and smaller, with less than ~5% by mass of contamination with larger RNAs.

Solid tissues or cells are disrupted by grinding or vortexing in BiooPure RNA Extraction Reagent. The resulting lysate is mixed with 1-bromo-3-chloropropane and centrifuged, which results in phase separation with the RNA partitioned in the upper aqueous phase. The aqueous phase is recovered and mixed with ethanol suitable for selective precipitation of large RNA, which is removed by solid phase extraction on a silica filter. The flow-through containing small RNA is recovered and mixed with additional ethanol to precipitate the small RNA, which is then purified by solid phase extraction onto a second silica filter. The filter is washed and the RNA eluted in RNase-free water containing a trace amount of EDTA.

Benefits
• Designed for NGS applications
• Fast and easy small RNA extraction
• Liquid nitrogen is not required for tissue disruption
• Offers high yields of small RNA
• Ideal for enrichment of microRNAs and other small RNAs from eukaryotic cells and tissues
• Validated with NEXTflex™ Sequencing Kits