Poly(A) RNA Selection Kit V1.5

The Poly(A) RNA Selection Module enables the rapid and highly specific enrichment of polyadenylated RNAs from total RNA samples.

Optimized V1.5 Upgrade

The upgrade V1.5 contains new oligo(dT) Magnetic Beads (MB), compared to the V1.0 kit. The beads and solutions included in the V1.5 kit offer improved performance.

Highly Specific for Poly(A) RNA

Other RNA species (rRNA and tRNA) do not contain poly(A) sequences and therefore will not bind to the oligo(dT) beads.

Poly(A) Figure 1

Figure 1. Bioanalyzer traces of Universal Human Reference Total RNA (UHRR, red trace) and poly(A) selected RNA from 5 µg UHRR (blue trace).

Various Downstream Applications

Isolated mRNA can directly be used for RNA-Seq library preparation (e.g., CORALL RNA-Seq Library Prep kits), SAGE, CAGE, cloning, microarrays, cDNA synthesis, and others.

Rapid Turnaround

Polyadenylated RNAs can be isolated from total RNA samples within about one hour.


Preparation of Beads and Denaturation of RNA
Step 1:
The total RNA Is briefly denatured and magnetic beads
are aliquoted and washed.
Hybridization of poly(A) RNA
Step 2:
The polyadenylated 3’ ends present in most mRNAs
are hybridized to oligodT beads.
Poly(A) RNA Enrichment
Step 3:
Any RNA without poly(A) stretches, such as rRNA
and tRNA,will not be captured by the oligodT beads
and will be washed away.
Poly(A) RNA Elution
Step 4:
The poly(A) RNA is eluted from the olidodT beads
by heating in water.


Frequently Asked Questions

Please find a list of the most frequently asked questions below. If you cannot find the answer to your question here or want to know more about our products, please contact

The oligodT consists of 30 Ts.

The poly(A) RNA Selection Module enriches for all RNAs that have a poly(A) tail, such as mRNA, but also mitochondrial rRNA, which can make up to 4 % of the isolated poly(A) RNA (=0.08% of the total RNA). Therefore, between 2 – 4 % of sequencing reads can correspond to rRNA when using various standard RNA-Seq library preps.

The Poly(A) RNA Selection Kit V1.5 features new oligo(dT) Magnetic Beads (MB) compared to the V1.0 kit. The V1.5 kit protocol has been carefully optimized to maximize poly(A) RNA recovery. Additional specific changes made in the V1.5 Kit and protocol include:

  • Updated Hybridization (HYB) and Bead Wash (BW) buffer compositions (for compatibility with new oligo(dT) Magnetic Beads).
  • Reduction of the Magnetic Bead volume to 2 µl (vs. 10 µl in V1.0), which can be used for total RNA inputs up to 5 µg.
Repeated rounds of poly(A) selection are not required due to the high specificity of the poly(A) selection. One poly(A) selection round is enough. Repeated rounds are often performed to remove unspecifically bound rRNA.
The amount of poly(A) RNA varies depending on the tissue and cell type the total RNA is derived from. Poly(A) RNA usually makes up 1-3 % of the total RNA.
Agilent’s Bioanalyzer and other electrophoresis platforms often try to detect 18S and 28S rRNA peaks at all costs. Therefore, highly abundant transcripts are sometimes wrongly identified as rRNA.

In the traces below you’ll see that the Bioanalyzer software classifies two peaks within the trace of a poly(A) selected sample as 18S and 28S rRNA (Figure A). However, the overlay with diluted total RNA clearly demonstrates that those peaks are not rRNA peaks (Figure B). Poly(A) selection was successful and no rRNA was left. This was also confirmed by NGS sequencing.


Figure A. Bioanalyzer trace after the Poly(A) RNA Selection using 5 µg of Universal Human Reference RNA (UHRR). The Bioanalyzer software wrongly classifies two peaks within the trace of the poly(A) selected sample as 18S and 28S rRNA.


Figure B. Overlay of Bioanalyzer traces before and after the Poly(A) RNA Selection. The overlay with diluted total RNA clearly demonstrates that those peaks identified as rRNA (in A) are not rRNA peaks.
Red trace: diluted total Universal Human Reference RNA (UHRR), intact (RIN 8.5). Blue trace: poly(A) RNA selected RNA.

Increasing the proportional bead volume to suit the input RNA amount will enable poly(A) selection from higher inputs. For recommendations based on your intended input please contact


pdf  User Guide – update 11.11.2020
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Material Safety Datasheets

MSDS Information can be found in the Documents page.

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