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QIAseq FastSelect RNA Removal Kits

For rapid rRNA and/or globin mRNA removal for RNA-seq library preparation
  • Compatible with QIAGEN, Illumina, NEB and KAPA RNA stranded library kits
  • High performance rRNA and/or globin removal in just 20 minutes
  • Only one pipetting step – combine removal reagent with RNA and incubate
  • No extra cleanup steps or protocol modifications
  • Customize to your experiments by species and targets

NEW!  QIAseq FastSelect –rRNA HMR Kits for rRNA and/or globin mRNA removal are 30% faster and more powerful than our previous version. Click here to view!

QIAseq FastSelect RNA Removal Kits use a novel method to remove highly abundant RNA that is of low scientific value from your RNA-seq libraries. Whether you are performing whole transcriptome analysis and want to remove cytoplasmic and mitochondrial rRNA or doing gene expression research from blood samples and require globin mRNA removal, QIAseq FastSelect Kits specifically remove the targeted RNA in a single pipetting step, without additional protocol changes or cleanup steps. 
Also look at QIAseq Stranded RNA Library Kits for robust RNA-seq library preparation.

Interested in customization of a QIAseq FastSelect RNA Removal Kit for your samples? Contact us!

Cat No./ID: 333180
QIAseq FastSelect RNA Removal Kit
134,541.03 kr
Go to GeneGlobe
Reagents for rRNA removal or globin mRNA removal for 24, 96 or 384 samples; human, mouse or rat
Cat No./ID: 333280
QIAseq FastSelect Multi-RNA Removal Kit
224,241.03 kr
Go to GeneGlobe
Reagents for rRNA and globin mRNA removal for 24, 96 or 384 samples; human, mouse or rat

QIAseq FastSelect RNA Removal Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


0
QIAseq FastSelect RNA removal workflow.
QIAseq FastSelect RNA Removal is a one-step rRNA and/or globin mRNA depletion solution. Simply add QIAseq FastSelect reagent (rRNA Removal and/or Globin Removal) to RNA sample, perform fragmentation (if required), stepwise cool the reaction from 75°C to 25°C for 20 minutes and then complete the remaining library preparation steps. QIAseq FastSelect works with or without RNA fragmentation, providing the flexibility to use FFPE or degraded RNA samples, or high-quality RNA as part of a standard RNA-seq library construction workflow.

QIAseq FastSelect has been tested with the QIAseq Stranded Total RNA Lib Kit (QIAGEN), TruSeq Stranded (Illumina), NEBNext Ultra II Directional (New England Biolabs) and KAPA RNA HyperPrep (KAPA Biosystems) library preparation kits. Specific requirements for fragmentation, as well as the duration of the fragmentation process, depend on the library preparation kit and sample type. Fragmentation is not required for QIAseq FastSelect performance.
1
QIAseq FastSelect vs. no treatment, Poly-A enrichment and Ribo-Zero Gold (% rRNA remaining).
Stranded transcriptome libraries were prepared from 100 ng aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the QIAseq Stranded Total RNA Lib Kit. UHRR samples were either: 1. Not treated, 2. Poly-A enriched, 3. rRNA-depleted with Ribo-Zero Gold or 4. rRNA-depleted using QIAseq FastSelect. Poly-A enrichment and Ribo-Zero Gold rRNA depletion were performed according to the manufacturers’ instructions. For QIAseq FastSelect rRNA depletion, during the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

A and B QIAseq FastSelect results in highly efficient removal of rRNA to levels observed with Poly-A enrichment. Average % rRNA remaining is provided in A and the individual replicates are plotted in B.
2
QIAseq FastSelect vs. no treatment, Poly-A enrichment and Ribo-Zero Gold (gene expression results).
Stranded transcriptome libraries were prepared from 100 ng aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the QIAseq Stranded Total RNA Lib Kit. UHRR samples were either: 1. Not treated, 2. Poly-A enriched, 3. rRNA-depleted with Ribo-Zero Gold or 4. rRNA-depleted using QIAseq FastSelect. Poly-A enrichment and Ribo-Zero Gold rRNA depletion were performed according to the manufacturers’ instructions. For QIAseq FastSelect rRNA depletion, during the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

A When QIAseq FastSelect is used for rRNA depletion, gene expression results are highly similar to results observed when using Ribo-Zero Gold (log2 normalized gene expression), demonstrating the efficacy of QIAseq FastSelect. B Gene expression results are highly similar between QIAseq FastSelect and untreated samples (no treatment), suggesting QIAseq FastSelect does not perturb the natural expression profiles of samples (targets with greater than 10 reads in both sample sets [n=3 each] have been plotted). C Gene expression profiles from Poly-A enriched samples are very different from untreated samples (no treatment), suggesting Poly-A enrichment perturbs the natural expression profile of samples (targets with greater than 10 reads in both sample sets [n=3] have been plotted).






3
QIAseq FastSelect performance with sequencing quality control (SEQC) samples (% rRNA remaining).
Stranded transcriptome libraries were prepared from 100 ng aliquots of the SEQC reference samples using the QIAseq Stranded Total RNA Lib Kit (Universal Human Reference RNA = UHRR [Agilent] and Human Brain Reference RNA = HBRR [Thermo Scientific]). QIAseq FastSelect was used for rRNA depletion. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

QIAseq FastSelect results in highly efficient removal of rRNA from the SEQC samples. Average % rRNA remaining is provided in A and the individual replicates are plotted in B.
4
QIAseq FastSelect performance with sequencing quality control (SEQC) samples (gene expression results).
Stranded transcriptome libraries were prepared from 100 ng aliquots of the SEQC reference samples using the QIAseq Stranded Total RNA Lib Kit (Universal Human Reference RNA = UHRR [Agilent] and Human Brain Reference RNA = HBRR [Thermo Scientific]). QIAseq FastSelect was used for rRNA depletion. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Three replicates were performed for each sample and gene expression results are highly reproducible (log2 normalized gene expression) when QIAseq FastSelect is used for rRNA depletion.
5
QIAseq FastSelect performance with sequencing quality control (SEQC) samples (principal component analysis).
Stranded transcriptome libraries were prepared from 100 ng aliquots of the SEQC reference samples using the QIAseq Stranded Total RNA Lib Kit (Universal Human Reference RNA = UHRR [Agilent] and Human Brain Reference RNA = HBRR [Thermo Scientific]). QIAseq FastSelect was used for rRNA depletion. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Principal component analysis (PCA) of expression profiles demonstrate that the replicate samples cluster as expected with each other and the four reference samples cluster as expected in relation to one another.
6
QIAseq FastSelect performance with sequencing quality control (SEQC) samples (Venn diagram).
Stranded transcriptome libraries were prepared from 100 ng aliquots of the SEQC reference samples using the QIAseq Stranded Total RNA Lib Kit (Universal Human Reference RNA = UHRR [Agilent] and Human Brain Reference RNA = HBRR [Thermo Scientific]). QIAseq FastSelect was used for rRNA depletion. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Venn diagram of differentially expressed genes demonstrates unique and overlapping expression profiles across the samples. The Venn diagram shows the number of genes with ≥2-fold change when FDR p-value is ≤0.05.
7
QIAseq Fast Select shows robust performance with RNA from FFPE samples (% rRNA remaining).
Total RNA was isolated from 5 µm normal and cancer lung FFPE sections using the miRNeasy FFPE Kit. Stranded transcriptome libraries were subsequently prepared from 100 ng aliquots using the QIAseq Stranded Total RNA Lib Kit. For rRNA depletion, QIAseq FastSelect or Ribo-Zero Gold was used. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.
QIAseq FastSelect: During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer, heated (fragmented) to 75°C for 2 min and stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed.
Ribo-Zero Gold: Prior to QIAseq library preparation, rRNA was removed from each RNA sample using Ribo-Zero Gold, which required over 2 hours to complete. Afterwards, QIAseq RNA library preparation was performed.
A Average % rRNA remaining. B Identical colors represent the same sample and replicates. QIAseq FastSelect results in highly efficient removal of rRNA from fragmented samples. Average % rRNA remaining is provided in A and plotted in B. Ribo-Zero Gold required substantially more amplification cycles than the QIAseq FastSelect libraries, suggesting some sample may be lost with Ribo-Zero Gold.





8
Highly reproducible gene expression data following rRNA depletion for RNA from FFPE samples.
Total RNA was isolated from 5 µm normal and cancer lung FFPE sections using the miRNeasy FFPE Kit. Stranded transcriptome libraries were subsequently prepared from 100 ng aliquots using the QIAseq Stranded Total RNA Lib Kit. For rRNA depletion, QIAseq FastSelect or Ribo-Zero Gold was used. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.
QIAseq FastSelect: During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer, heated (fragmented) to 75°C for 2 min and stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed.
Ribo-Zero Gold: Prior to QIAseq library preparation, rRNA was removed from each RNA sample using Ribo-Zero Gold, which required over 2 hours to complete. Afterwards, QIAseq RNA library preparation was performed.
Performing rRNA depletion using QIAseq FastSelect achieved highly reproducible gene expression results (log2 normalized gene expression) for A normal FFPE RNA and B cancer FFPE RNA. Two replicates were performed for each sample type.
9
QIAseq FastSelect provides robust performance with FFPE standards (% rRNA remaining).
Total RNA was isolated from FFPE reference standards (Fusion RNA Multiplex Positive and Negative Controls [Horizon™]) using the miRNeasy FFPE Kit. Stranded transcriptome libraries were subsequently prepared from 100 ng aliquots using the QIAseq Stranded Total RNA Lib Kit. For rRNA depletion, QIAseq FastSelect or Ribo-Zero Gold was used. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.
QIAseq FastSelect: During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed.
Ribo-Zero Gold: Prior to QIAseq library preparation, rRNA was removed from each RNA sample using Ribo-Zero Gold, which required over 2 hours to complete. Afterwards, QIAseq RNA library preparation was performed.
A Average % rRNA remaining. B Identical colors represent the same sample and replicates. Average % rRNA remaining is provided in A and the individual replicates are plotted in B. QIAseq FastSelect demonstrated highly efficient removal of rRNA from FFPE standards. Ribo-Zero Gold required substantially more amplification cycles than the QIAseq FastSelect libraries, suggesting some sample may be lost with Ribo-Zero Gold.










10
QIAseq FastSelect provides robust performance with FFPE standards (gene expression results).
Total RNA was isolated from FFPE reference standards (Fusion RNA Multiplex Positive and Negative Controls [Horizon]) using the miRNeasy FFPE Kit. Stranded transcriptome libraries were subsequently prepared from 100 ng aliquots using the QIAseq Stranded Total RNA Lib Kit. For rRNA depletion, QIAseq FastSelect or Ribo-Zero Gold was used. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.
QIAseq FastSelect: During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 75°C for 2 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed.
Ribo-Zero Gold: Prior to QIAseq library preparation, rRNA was removed from each RNA sample using Ribo-Zero Gold, which required over 2 hours to complete. Afterwards, QIAseq RNA library preparation was performed.
Performing rRNA depletion using QIAseq FastSelect achieved highly reproducible gene expression results (log2 normalized gene expression) for A Fusion RNA Multiplex Positive FFPE and B Fusion RNA Multiplex Negative FFPE. Two replicates were performed for each sample type.














11
QIAseq FastSelect provides robust performance with RNA from tissue samples (% rRNA remaining).
Stranded transcriptome libraries were prepared from 100 ng aliquots of human tissue RNA samples using the QIAseq Stranded Total RNA Lib Kit. QIAseq FastSelect was used for rRNA depletion. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

A Average % rRNA remaining. B Tissue replicates (n=3). QIAseq FastSelect results in highly efficient removal of rRNA from RNA samples extracted from various human tissues. Average % rRNA remaining is provided in A and the individual tissue replicates are plotted in B.
12
QIAseq FastSelect provides robust performance with RNA from tissue samples (gene expression results).
Stranded transcriptome libraries were prepared from 100 ng aliquots of human tissue RNA samples using the QIAseq Stranded Total RNA Lib Kit. QIAseq FastSelect was used for rRNA depletion. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Performing rRNA depletion using QIAseq FastSelect achieved highly reproducible gene expression results (log2 normalized gene expression) for RNA samples extracted from various human tissues. Three replicates were performed for each sample.
13
QIAseq FastSelect provides robust performance with RNA from cell lines (% rRNA remaining).
Stranded transcriptome libraries were prepared from 100 ng and 1µg aliquots of HeLa S3 and HCT 116 total RNA samples using the QIAseq Stranded Total RNA Lib Kit. QIAseq FastSelect was used for rRNA depletion. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a MiSeq (Illumina) and data analysis was performed using CLC Biomedical Workbench.

A Average % rRNA remaining. B Blue = QIAseq FastSelect and red = no treatment. QIAseq FastSelect results in highly efficient removal of rRNA compared to untreated samples. Average % rRNA remaining is provided in A and plotted in B.
14
QIAseq FastSelect provides robust performance with RNA from cell lines (gene expression results).
Stranded transcriptome libraries were prepared from 100 ng and 1µg aliquots of A HeLa S3 and B HCT 116 total RNA samples using the QIAseq Stranded Total RNA Lib Kit. QIAseq FastSelect was used for rRNA depletion. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a MiSeq (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Performing rRNA depletion using QIAseq FastSelect achieved highly reproducible gene expression results (log2 normalized gene expression) for RNA samples extracted from cell lines when comparing 100 ng (X-axis) to 1 µg (Y-axis). Two replicates were performed for each sample.
15
QIAseq FastSelect provides robust lot-to-lot reproducibility for RNA depletion (% rRNA remaining).
Stranded transcriptome libraries were prepared from 100 ng aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the QIAseq Stranded Total RNA Lib Kit. For rRNA depletion, two different lots of QIAseq FastSelect were used. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a MiSeq (Illumina) and data analysis was performed using CLC Biomedical Workbench.

A Average % rRNA remaining and % protein coding reads. B Blue = QIAseq FastSelect lot 1 and red = QIAseq FastSelect lot 2. QIAseq FastSelect results in highly efficient removal of rRNA regardless of kit lot. Average % rRNA remaining is provided in A and plotted in B.
16
QIAseq FastSelect provides robust lot-to-lot reproducibility for RNA depletion (gene expression results).
Stranded transcriptome libraries were prepared from 100 ng aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the QIAseq Stranded Total RNA Lib Kit. For rRNA depletion, two different lots of QIAseq FastSelect were used. During the QIAseq library preparation, rRNA removal reagent was combined with each RNA sample and 5x RT Buffer. Next, the samples were heated (fragmented) at 95°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed. Sequencing was performed on a MiSeq (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Performing rRNA depletion using QIAseq FastSelect achieved highly reproducible gene expression results (log2 normalized gene expression) when using different kit lots. Three replicates were performed for each sample.
17
QIAseq FastSelect integrated into Illumina TruSeq workflow (performance).
Stranded transcriptome libraries were prepared from 100 ng and 1 µg aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the Illumina TruSeq Stranded mRNA Library Prep kit (mRNA enrichment was not performed). For rRNA depletion, QIAseq FastSelect was used. During the TruSeq library preparation, rRNA removal reagent was combined with each RNA sample and FPF (from the TruSeq kit). Next, the samples were heated (fragmented) at 94°C for 8 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed according to the TruSeq recommendations. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

A Average % rRNA remaining and % protein coding reads. B rRNA remaining is shown when no depletion is performed (no treatment) and when depletion is performed using QIAseq FastSelect (blue = 100 ng and red = 1 µg). QIAseq FastSelect results in highly efficient removal of rRNA. Average % rRNA remaining is provided in A and plotted in B.
18
QIAseq FastSelect integrated into Illumina TruSeq workflow (gene expression results).
Stranded transcriptome libraries were prepared from A 100 ng and B 1 µg aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the Illumina TruSeq Stranded mRNA Library Prep kit (mRNA enrichment was not performed). For rRNA depletion, QIAseq FastSelect was used. During the TruSeq library preparation, rRNA removal reagent was combined with each RNA sample and FPF (from the TruSeq kit). Next, the samples were heated (fragmented) at 94°C for 8 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed according to the TruSeq recommendations. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Performing rRNA depletion using QIAseq FastSelect achieved highly reproducible gene expression results (log2 normalized gene expression). Two replicates were performed for each sample.
19
QIAseq FastSelect integrated into NEBNext Ultra II Directional RNA Library workflow (performance).
Stranded transcriptome libraries were prepared from 5 ng and 1 µg aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the NEBNext Ultra II Directional RNA Library Prep kit (New England Biolabs). For rRNA depletion, QIAseq FastSelect was used. During the NEBNext library preparation, rRNA removal reagent was combined with each RNA sample, 5x NEBNext First Strand Synthesis Reaction Buffer (from the NEBNext kit) and random primers (from the NEBNext kit). Next, the samples were heated (fragmented) at 94°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed according to the NEBNext recommendations. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

A Average % rRNA remaining and % protein coding reads. B rRNA remaining is shown when no depletion is performed (no treatment) and when depletion is performed using QIAseq FastSelect (blue = 5 ng and red = 1 µg). QIAseq FastSelect results in highly efficient removal of rRNA. Average % rRNA remaining is provided in A and plotted in B.
20
QIAseq FastSelect integrated into NEBNext Ultra II Directional RNA Library workflow (gene expression results).
Stranded transcriptome libraries were prepared from A 5 ng and B 1 µg aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the NEBNext Ultra II Directional RNA Library Prep kit (New England Biolabs). For rRNA depletion, QIAseq FastSelect was used. During the NEBNext library preparation, rRNA removal reagent was combined with each RNA sample, 5x NEBNext First Strand Synthesis Reaction Buffer (from the NEBNext kit) and random primers (from the NEBNext kit). Next, the samples were heated (fragmented) at 94°C for 15 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed according to the NEBNext recommendations. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Performing rRNA depletion using QIAseq FastSelect achieved highly reproducible gene expression results (log2 normalized gene expression). Two replicates were performed for each sample.
21
QIAseq FastSelect integrated into KAPA RNA HyperPrep workflow (performance).
Stranded transcriptome libraries were prepared from 25 ng and 1 µg aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the KAPA RNA HyperPrep Kit (KAPA Biosystems). For rRNA depletion, QIAseq FastSelect was used. During the KAPA RNA HyperPrep workflow, rRNA removal reagent was combined with each RNA sample and Fragment, Primer and 2x Elute Buffer (from the KAPA RNA HyperPrep kit). Next, the samples were heated (fragmented) at 94°C for 8 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed according to the KAPA RNA HyperPrep recommendations. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

A Average % rRNA remaining and % protein coding reads. B rRNA remaining is shown when depletion is performed using QIAseq FastSelect (blue = 25 ng and red = 1 µg). QIAseq FastSelect results in highly efficient removal of rRNA. Average % rRNA remaining is provided in A and plotted in B.
22
QIAseq FastSelect integrated into KAPA RNA HyperPrep workflow (gene expression results).
Stranded transcriptome libraries were prepared from A 25 ng and B 1 µg aliquots of Universal Human Reference RNA (UHRR) (Agilent) using the KAPA RNA HyperPrep Kit (KAPA Biosystems). For rRNA depletion, QIAseq FastSelect was used. During the KAPA RNA HyperPrep workflow, rRNA removal reagent was combined with each RNA sample and Fragment, Primer and 2x Elute Buffer (from the KAPA RNA HyperPrep kit). Next, the samples were heated (fragmented) at 94°C for 8 min and then stepwise cooled to 25°C for 20 minutes. Afterwards, the remaining library preparation steps were performed according to the KAPA RNA HyperPrep recommendations. Sequencing was performed on a NextSeq 550 (Illumina) and data analysis was performed using CLC Biomedical Workbench.

Performing rRNA depletion using QIAseq FastSelect achieved highly reproducible gene expression results (log2 normalized gene expression). Two replicates were performed for each sample.
Performance

QIAseq FastSelect RNA Removal Kits result in highly efficient removal of rRNA to levels observed with Poly-A enrichment. Using sequencing quality control samples (SEQC), QIAseq FastSelect demonstrated high performance rRNA removal and highly reproducible gene expression results.
See Figures:
QIAseq FastSelect vs. no treatment, Poly-A enrichment and Ribo-Zero Gold (% rRNA remaining)
QIAseq FastSelect vs. no treatment, Poly-A enrichment and Ribo-Zero Gold (gene expression results)
QIAseq FastSelect performance with sequencing quality control (SEQC) samples (% rRNA remaining)
QIAseq FastSelect performance with sequencing quality control (SEQC) samples (gene expression results)
QIAseq FastSelect performance with sequencing quality control (SEQC) samples (principal component analysis)
QIAseq FastSelect performance with sequencing quality control (SEQC) samples (Venn diagram)

Robust performance with a variety of sample types

QIAseq FastSelect provides reliable rRNA removal and reproducible gene expression results with RNA extracted from various sample types, including FFPE, human tissues and cell lines.
See Figures:
QIAseq Fast Select shows robust performance with RNA from FFPE samples (% rRNA remaining)
Highly reproducible gene expression data following rRNA depletion for RNA from FFPE samples
QIAseq FastSelect provides robust performance with FFPE standards (% rRNA remaining)
QIAseq FastSelect provides robust performance with FFPE standards (gene expression results)
QIAseq FastSelect provides robust performance with RNA from tissue samples (% rRNA remaining)
QIAseq FastSelect provides robust performance with RNA from tissue samples (gene expression results)
QIAseq FastSelect provides robust performance with RNA from cell lines (% rRNA remaining)
QIAseq FastSelect provides robust performance with RNA from cell lines (gene expression results)

Integrates with existing stranded library construction workflows

Requiring just a single, inline pipetting step, QIAseq FastSelect is fully compatible with Illumina, NEB and KAPA RNA stranded library kits.
See Figures:
QIAseq FastSelect integrated into Illumina TruSeq workflow (performance)
QIAseq FastSelect integrated into Illumina TruSeq workflow (gene expression results)
QIAseq FastSelect integrated into NEBNext Ultra II Directional RNA Library workflow (performance)
QIAseq FastSelect integrated into NEBNext Ultra II Directional RNA Library workflow (gene expression results)
QIAseq FastSelect integrated into KAPA RNA HyperPrep workflow (performance)
QIAseq FastSelect integrated into KAPA RNA HyperPrep workflow (gene expression results)


Superior performance, faster workflow

QIAseq FastSelect is considerably faster than other methods and provides superior performance.
See Figures:
QIAseq Fast Select shows robust performance with RNA from FFPE samples (% rRNA remaining)
Highly reproducible gene expression data following rRNA depletion for RNA from FFPE samples
QIAseq FastSelect provides robust performance with FFPE standards (% rRNA remaining)
QIAseq FastSelect provides robust performance with FFPE standards (gene expression results)
QIAseq FastSelect provides robust lot-to-lot reproducibility for RNA depletion (% rRNA remaining)
QIAseq FastSelect provides robust lot-to-lot reproducibility for RNA depletion (gene expression results)

Principle
Removing highly expressed, but biologically unimportant RNA transcripts makes NGS more efficient and enables higher sample throughput with higher sensitivity. Furthermore, RNA from FFPE samples and degraded RNA samples are particularly challenging for RNA removal and can result in suboptimal performance.

QIAseq FastSelect is designed for quick, efficient removal of cytoplasmic and mitochondrial rRNA and/or globin mRNA from total RNA during NGS RNA library preparation. QIAseq FastSelect seamlessly integrates with your existing RNA stranded library preparation workflow for RNA removal in a single, 20-minute inline step. Prior to RNA heat fragmentation (which is optional and dependent upon the library preparation kit and sample type), QIAseq FastSelect removal reagent is directly combined with total RNA and the library preparation-specific buffers. After fragmentation, the reaction temperature is step-wise cooled to room temperature and the remaining library preparation steps are completed. There is no need to perform any type of enrichment on the total RNA samples. QIAseq FastSelect RNA Removal Kits can accommodate RNA amounts ranging from as little as 1 ng up to 1 µg with consistently high performance. QIAseq FastSelect can be used with RNA from fresh samples, as well as RNA from FFPE (formalin-fixed paraffin-embedded) samples or degraded RNA, resulting in reliable rRNA removal and high reproducibility in downstream applications.
Procedure
Most RNA removal or depletion strategies associated with RNA-seq library construction are sample pre-treatment strategies involving hybrid-capture or enzymatic removal of unwanted RNA. Our unique QIAseq FastSelect procedure is compatible with QIAGEN, Illumina, KAPA and NEB stranded library preparation kits and provides complete rRNA removal in a single, 20-minute inline step (Figure QIAseq FastSelect RNA removal workflow). This is dramatically faster than competing RNA depletion kits, which require pre-treatment protocols involving more than 25 steps and 2 hours to complete.

Simply add QIAseq FastSelect reagent (rRNA Removal and/or Globin Removal) to the RNA sample, perform fragmentation (if required), stepwise cool the reaction from 75°C to 25°C for 20 minutes and then complete the remaining library preparation steps. QIAseq FastSelect works with or without RNA fragmentation, providing the flexibility to use RNA from FFPE samples or degraded RNA samples, or high-quality RNA as part of a standard RNA-seq library construction workflow.
Applications

QIAseq FastSelect delivers rapid, reliable RNA removal for FFPE and fresh sample RNA sources. QIAseq FastSelect RNA Removal Kits are available in a variety of different formats and sizes to suit your specific applications. QIAGEN offers kits for removing human, mouse and rat RNA, as well as a customizable product (coming soon) designed for the species and RNA of your choice.

Wide array of QIAseq FastSelect products
Product Species available Targets Number of samples
QIAseq FastSelect RNA Removal Kit Human, mouse and rat rRNA or globin mRNA 24, 96 or 384
QIAseq FastSelect Multi-RNA Removal Kit Human, mouse and rat rRNAand globin mRNA 24, 96 or 384
QIAseq FastSelect Custom RNA Removal Kit (coming soon) Your choice Any RNA 1156
 

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