Invitrogen™ RiboMinus™ Transcriptome Isolation Kit, human/mouse

Catalog No.	Quantity
K155002	6 preps

Catalog No. K155002 Supplier Invitrogen™ Supplier No. K155002

Description

Includes

The 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.

RiboMinus™ Transcriptome Isolation Kits are novel purification systems that enrich the whole spectrum of RNA transcripts by depleting large ribosomal RNA molecules in a total RNA sample.

RiboMinus™ Transcriptome Isolation Kits are novel purification systems that enrich the whole spectrum of RNA transcripts by depleting large ribosomal RNA molecules in a total RNA sample. These kits use a 5μ biotin-labeled, Locked Nucleic Acid (LNA) probe specific for large rRNA to remove 95 to 98% of rRNA molecules from human, mouse, bacteria, or yeast total RNA. This removal increases the representation of RNA transcript species, including non-poly(A) tailed, pre-processed RNA, small tRNAs, siRNA, snRNA, microRNA, fragmented mRNA, and other regulatory RNAs that cannot be purified via poly(A) selection methods.

Purified RNA is concentrated and suitable for numerous downstream applications, including microarray analysis, library construction, and qRT-PCR.

• Comprehensive analysis of the whole transcriptome without contamination or interference from rRNA
• More complete collection of transcribed elements of the genome—no reliance on poly(A) purification methods
• Cleaner, clearer microarray analysis due to higher signal-to-background ratios
• Not high-throughput compatible (Manual)

Specifications

• Content And Storage: The 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.
• Isolation Technology: Magnetic Bead
• Purification Time: 1.25 hr.
• Sample Type: Total RNA
• Final Product Type: Transcriptome RNA
• High-throughput Compatibility: Not High-throughput Compatible (Manual)
• For Use With (Application): Microarray Analysis, Real-Time Quantitative PCR (qPCR), Reverse Transcriptase PCR (RT-PCR), Northern Blotting, cDNA Library Construction
• Product Line: RiboMinus
• Quantity: 6 preps
• Starting Material Amount: Up to 10 μg total RNA
• Target: Transcriptome RNA
• System Type: RiboMinus
• Yield: 1 μg
• Form: Magnetic Bead

Frequently Asked Questions (FAQs)

I used the Ribominus Human/Mouse Transcriptome Isolation Kit to deplete large ribosomal RNA from human total RNA but I am seeing genomic DNA contamination. Why is this?

This is most likely due to contamination of the total RNA sample with genomic DNA. Before the RiboMinus Kit procedure, we recommend treating the total RNA sample with DNase I to remove any genomic DNA contamination.

What is the best way to analyze RNA that I've isolated using a RiboMinus kit?

The purified RNA is easily quantitated using UV absorbance at 260 nm or a Quant-iT RNA Assay Kit. To verify rRNA depletion, electrophorese your sample on an agarose gel or use an Agilent 2100 Bioanalyzer. Agarose gel electrophoresis should show depletion of 18S and 28S rRNA bands compared to a control sample. Absence of contaminating DNA and RNA degradation may also be confirmed by agarose gel electrophoresis. The efficiency of rRNA depletion in the purified RNA, RNA degradation, and RNA concentration can also be analyzed using a bioanalyzer.

What species have you tested with your RiboMinus Eukaryote Kit for RNA-Seq?

With regard to species specificity, the following organisms have 0 mismatches for the LSU/SSU probes (LSU (28S) / SSU (18S) Probe perfect matches). This means the probes will remove the 18S and 28S sequences from the following species.

- Species from the Land: Human, mouse, rat, frog, rabbit, cow, pig, chicken, Drosophila,* Caenorhabditis elegans, midge, mosquito, yeast (Saccharomyces cerevisiae, Schizosaccharomyces pombe)
- Species from the Sea: Sea squirt, coelacanth, lancelet, eel, salmon, sturgeon, ratfish, lamprey, brown plankton, zooplankton
- Plants: Arabidopsis thaliana, Brassica napus, rice, tomato, Humulus lupulus, Zea mays, wheat, soybean, pine, aspen

* The 28S and 18S probes in this kit respectively align 100% to the Drosophila 18S and 28S rRNA sequence. However, there are reports suggesting Drosophila (insect) 28S rRNA split into 2 smaller fragments co-migrating with 18S. The exact split site is not identified yet. But based on customer feedback, this kit does not remove the split 28S rRNA completely. We have not tested Drosophila samples with this kit.

The 5S and 5.8S sequences are more divergent, and thus the best probes possible have reduced species breadth:
- 5S: zero to 1 mismatch: human, mouse, rat, frog, chicken, Drosophila, C. elegans, A. thaliana, Z. mays
- 5.8S: zero mismatches: human, mouse, rat

For Research Use Only. Not for use in diagnostic procedures.