Globin mRNA and rRNA are highly abundant in blood samples, and their efficient and specific removal is desirable in order to reveal the biological significance of less abundant transcripts.
The NEBNext® Globin & rRNA Depletion Kit (Human/Mouse/Rat) employs the NEBNext RNase H-based RNA depletion workflow to deplete globin mRNA, cytoplasmic rRNA and mitochondrial rRNA with human, mouse and rat samples, and is effective with intact and degraded RNA.
The kit is also available with RNAClean® beads.
The great majority of RNA in blood samples is comprised of cytoplasmic and mitochondrial ribosomal RNAs (rRNA) and globin mRNA. These highly abundant RNA species can conceal the biological significance of less abundant transcripts, and so their efficient and specific removal is desirable.
The NEBNext® Globin & rRNA Depletion Kit (Human/Mouse/Rat) employs the NEBNext RNase H-based RNA depletion workflow to deplete adult, embryonic and fetal globin mRNA (HBA1/2, HBB, HBD, HBM, HBG1/2, HBE1, HBQ1 and HBZ), cytoplasmic rRNA (5S, 5.8S, 18S, 28S, ITS and ETS) and mitochondrial rRNA (12S and 16S).
The kit is effective with human, mouse and rat total RNA preparations, both intact and degraded. The resulting depleted RNA is suitable for RNA-seq, random-primed cDNA synthesis, or other downstream RNA analysis.
This kit can also be used following poly(A) mRNA enrichment (e.g., using the NEBNext poly(A) mRNA Magnetic Isolation Module NEB #E7490), so that the final depleted RNA contains only mRNA of interest and no non-coding RNA.
The kit is also available with RNAClean® beads.
Features
For use with NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors RNA Set 1) (NEB #E7416), refer to the Protocols tab for UMI Adaptors-specific guidance.
The following reagents are supplied with this product:
NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
E7750S | -20 |
NEBNext® DNase I | E7753-2VIAL | -20 | 1 x 0.015 ml | 20,000 units/ml | |
NEBNext® Thermostable RNase H | E7752-2VIAL | -20 | 1 x 0.012 ml | 5,000 units/ml | |
NEBNext® Globin and rRNA Depletion Solution | E7751-2VIAL | -20 | 1 x 0.018 ml | 1 X | |
RNase H Reaction Buffer | E6312-2VIAL | -20 | 1 x 0.012 ml | ||
DNase I Reaction Buffer | E6315-2VIAL | -20 | 1 x 0.03 ml | ||
Nuclease-free Water | E6317-2VIAL | -20 | 1 x 0.4 ml | ||
NEBNext Probe Hybridization Buffer | E6314-2VIAL | -20 | 1 x 0.012 ml |
E7750L | -20 |
NEBNext® DNase I | E7753-3VIAL | -20 | 1 x 0.06 ml | 20,000 units/ml | |
NEBNext® Thermostable RNase H | E7752-3VIAL | -20 | 1 x 0.048 ml | 5,000 units/ml | |
NEBNext® Globin and rRNA Depletion Solution | E7751-3VIAL | -20 | 1 x 0.072 ml | 1 X | |
RNase H Reaction Buffer | E6312-3VIAL | -20 | 1 x 0.048 ml | ||
DNase I Reaction Buffer | E6315-3VIAL | -20 | 1 x 0.12 ml | ||
Nuclease-free Water | E6317-3VIAL | -20 | 1 x 1.5 ml | ||
NEBNext Probe Hybridization Buffer | E6314-3VIAL | -20 | 1 x 0.048 ml |
E7750X | -20 |
NEBNext® DNase I | E7753-4VIAL | -20 | 1 x 0.24 ml | 20,000 units/ml | |
NEBNext® Thermostable RNase H | E7752-4VIAL | -20 | 1 x 0.192 ml | 5,000 units/ml | |
NEBNext® Globin and rRNA Depletion Solution | E7751-4VIAL | -20 | 1 x 0.288 ml | 1 X | |
RNase H Reaction Buffer | E6312-4VIAL | -20 | 1 x 0.192 ml | ||
DNase I Reaction Buffer | E6315-4VIAL | -20 | 1 x 0.48 ml | ||
Nuclease-free Water | E6317-4VIAL | -20 | 1 x 6 ml | ||
NEBNext Probe Hybridization Buffer | E6314-4VIAL | -20 | 1 x 0.192 ml |
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