Highly efficient depletion of 16S, 23S, and 5S rRNA from a variety of species The RiboMinus Bacteria 2.0 Transcriptome Isolation Kit is a complete and automatable kit that enables efficient and selective removal of bacterial and archaebacterial ribosomal RNA (rRNA) from intact total RNA (100 ng to 5 μg) in less than two hours.
Key features of the RiboMinus Bacteria 2.0 kit: • Efficient—the included RiboMinus Pan-Prokaryote Probe Mix has consistently shown less than 10% of reads mapping to 16S and 23S rRNA reads • Wide range of targets—designed with >150 probe designs targeting 16S, 23S, and 5S rRNA targets from >76 representative species of bacteria and archaebacteria from common phyla and sub-phyla • Convenient—Dynabeads MyOne Streptavidin C1 magnetic beads and bead cleanup module and protocols included for Kingfisher instruments • Flexible— accommodates mass inputs from 100 ng to 5 µg of total RNA; works with wide range of total RNA samples with RIN values from 5.0–10.0 • High detection sensitivity—increases the number of sequencing reads mapping to protein-coding transcripts compared to previous version • Suitable for metatranscriptomics—deplete rRNA of many microbial species in metagenomic samples
Target rRNA from panorama of bacteria and archaebacteria The RiboMinus Bacteria 2.0 kit is based on hybridization-capture technology and includes a new RiboMinus Pan-Prokaryote Probe Mix, which results in a 4-fold decrease of ribosomal reads remaining and shows significant improvement over the previous bacterial probe mix. The new probe mix consists of over 150 3’-biotin-labeled ssDNA oligonucleotiode designs. These designs were created using highly conserved regions of rRNA across 76 pan-prokaryote species (see table below) and tested with several species (see compatibility list below).
Dynabeads MyOne Streptavidin C1 cleanup kit included Once the probe mix has selectively hybridized to the target rRNA transcripts, removal of the probe-rRNA complex is completed by capture on Dynabeads MyOne Streptavidin C1 magnetic beads, leaving behind rRNA-depleted RNA. The RiboMinus Bacteria 2.0 kit includes a magnetic bead-based cleanup module for concentration of the rRNA-depleted RNA, enabling easier handling, scalability, and automation on KingFisher instruments. The resulting rRNA-depleted RNA (RiboMinus RNA) is suitable for whole-transcriptome analysis by RNA-Seq or other downstream applications.
Compatiblity The RiboMinus Pan-Prokaryote Probe Mix has been shown to be compatible with the following tested species: Acinetobacter baumannii Bacteroides thetaiotaomicron Bifidobacterium longum subsp. infantis. Campylobacter jejuni Clostridioides difficile Escherichia coli Fusobacterium nucleatum Helicobacter pylori Janthinobacterium sp. Moorella thermoacetica Pseudomonas putida Rickettsia rickettsia Salmonella typhimurium Pseudomonas aeruginosa Salmonella enterica Staphylococcus aureus Thermoanaerobacter kivui Vibrio cholerae Vibrio fisheri
The RiboMinus Pan-Prokaryote Probe Mix has been shown to be compatible with the following species based on sequence homology: Corynebacterium Enterococcus faecalis Gluconobacter Myxococcus xanthus Streptomyces coelicoflavus