Invitrogen™

RiboMinus™ Bacteria 2.0 Transcriptome Isolation Kit

Catalog number:  A47335

Related applications:

RNA Extraction

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Description

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

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Specifications

Final Product Type: Transcriptome RNA
For Use With (Application): Next Generation Sequencing, Microarray Analysis, Real-Time Quantitative PCR (qPCR), Reverse Transcriptase PCR (RT-PCR), cDNA Library Construction
Isolation Technology: Magnetic Bead
No. of Reactions: 12 Reactions
Purification Target: Transcriptome RNA
Sample Type: Total RNA (Bacterial)
Format: Magnetic Bead
Quantity: 1
High-throughput Compatibility: Not High-throughput Compatible (Manual)
Starting Material Amount: 100 ng to 5 ug of total RNA
System Type: RiboMinus
Product Line: RiboMinus™

Contents & storage

RiboMinus Isolation kits include RiboMinus Magnetic Beads, RiboMinus human/mouse, bacteria, or yeast probe, and Hybridization Buffer. The optional RiboMinus Concentration Module contains spin cartridges, wash tubes, recovery tubes, Binding Buffer, Wash Buffer, and RNase-free (DEPC-treated) water. Store the isolation modules at +4°C and the concentration module at room temperature. All reagents are guaranteed stable for 6 months when properly stored.