Sample Preparation |
rRNA Removal
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The new Ribo-Zero™ rRNA Removal Kit (Human/Mouse/Rat) Kit removes more ribosomal RNA (rRNA) from
intact and partially degraded human, mouse, and rat total RNA preparations
than any other method.
Benefits
- Removes >99.9% of 28S, 18S and 5.8S rRNA, and >95% of 5S rRNA from intact
and partially degraded human, mouse, and rat total RNA preparations.
- Single-pass process yields RNA sufficiently depleted of rRNA for RNA-Seq,
poly(A)-tailing, random-primed cDNA synthesis, and other applications.
- Recovers mRNAs, as well as other non-rRNAs that lack a poly(A) tail.
- Recovers both the 5´- and 3´-end fragments of mRNAs in a partially
degraded RNA sample.
| Figure 1. Overview of the Ribo-Zero™ rRNA Removal Kit (Human/Mouse/Rat)
process. |
Table 1. qRT-PCR analysis of Ribo-Zero™ rRNA Removal Kit (Human/Mouse/Rat)
treated RNA preparations. Intact and partially degraded HeLa total RNA
were treated using the Ribo-Zero kit. The amount of rRNA was then determined
by random-primed reverse transcription-qPCR using PCR primers to multiple
regions of each rRNA. Results of several experiments were averaged.
| Intact HeLa total RNA |
>99.9% |
>99.9% |
>99.9% |
96.8% |
| Partially degraded HeLa total RNA |
>99.9% |
>99.9% |
>99.9% |
97.8% |
Table 2. The Ribo-Zero™ rRNA Removal Kit (Human/Mouse/Rat) significantly
improves RNA-Seq results. Intact and partially degraded Universal Human
Reference RNA (UHRR) was treated with either the Ribo- Zero Kit or a competitive
kit. The rRNA-depleted RNAs were then used to prepare cDNA libraries that
were sequenced on an Illumina® GAII sequencer.
| Intact Human Reference RNA |
Ribo-Zero™ Kit |
1.4% |
| Intact Human Reference RNA |
Competitive Kit |
18.4% |
| Partially Degraded Human Reference RNA |
Ribo-Zero™ Kit |
2.1% |
| Partially Degraded Human Reference RNA |
Competitive Kit |
63.3% |
 |
Figure 2 (click to enlarge). Agilent Bioanalyzer analysis of human
Brain Total RNA before and after treatment with the Ribo-Zero™ rRNA
Removal Kit (Human/ Mouse/Rat). Two-hundred fifty nanograms of Brain
Total RNA (red) and the corresponding amount of Ribo-Zero-treated Brain
RNA (blue) were analyzed using an Agilent RNA 6000 Nano Kit. |
A |
 |
B |
C |
| Figure 3. Consistent gene expression data from Ribo-Zero™-treated
RNA. cDNA libraries were prepared and sequenced as described in Table
2, using Universal Human Reference RNA (UHRR). A) Number of genes detected
in libraries prepared from intact or partially degraded, rRNA-depleted
UHRR. B) Correlation of expression levels for intact RNA. C) Correlation
of expression levels for partially fragmented RNA. |
A |
B |
| Figure 4. RNA-Seq libraries prepared from Ribo-Zero™-treated
samples show very high correlation of differential gene expression compared
to MAQC qPCR data. RNA-Seq libraries were prepared by a random-primed
cDNA synthesis method from intact (A) or fragmented (B) Universal Human
Reference RNA (UHRR) and Human Brain Reference RNA (BrRR) total RNA samples,
after rRNA removal using the Ribo-Zero Kit. |
Ribo-Zero™ rRNA Removal Kit (Gram-Negative Bacteria)
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