QIAseq FastSelect –rRNA Yeast Kits

For rapid rRNA removal for RNA-seq library preparation from yeast samples

Features

  • Compatible with QIAGEN, Illumina, NEB and KAPA stranded RNA-seq library kits
  • Single reagent for yeast cells and spores
  • High-performance cytoplasmic and mitochondrial yeast rRNA removal in just 14 minutes
  • Only one pipetting step – combine QIAseq FastSelect reagent with RNA and incubate
  • No extra cleanup steps or NGS library protocol changes
undefined

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

QIAseq FastSelect –rRNA Yeast Kit (24)

Cat. No. / ID: 334215

Includes 3 tubes of QIAseq FastSelect reagent for cytoplasmic and mitochondrial rRNA removal; sufficient for 24 reactions from yeast samples
Samples
24
96
384
QIAseq FastSelect –rRNA Yeast Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

QIAseq FastSelect –rRNA Yeast Kits use a novel method to remove highly abundant RNA that is of low scientific value from your RNA-seq libraries. Researchers using RNA-seq for whole transcriptome analysis can use QIAseq FastSelect –rRNA Yeast Kits to remove cytoplasmic (35S [made up of 25S, 18S and 5.8S], 25S, 18S, 5.8S and 5S) and mitochondrial (21S, 15S, ACI60_gr01 [large subunit ribosomal RNA] and ACI60_gr02 [small subunit ribosomal RNA]) rRNA.

We recommend using QIAseq Stranded RNA Library Kits for robust strand-specific RNA-seq library preparation for both high-quality and highly fragmented RNA.

Design your own custom QIAseq FastSelect pools to remove any RNAs you wish from your RNA-seq library – take a look at our QIAseq FastSelect Custom RNA Removal Kits.

Want to try the QIAseq FastSelect –rRNA Yeast Kit for the first time? Request a trial kit to evaluate.

Performance

See figures

Principle

Removing highly expressed, but biologically unimportant yeast RNA transcripts before your RNA-seq library preparation protocol makes NGS more efficient and enables higher sample throughput with higher sensitivity. Furthermore, removal of unwanted RNA species from fragmented RNA samples is particularly challenging and can result in suboptimal performance.

QIAseq FastSelect Yeast Kits are designed for quick, efficient yeast rRNA removal from cytoplasmic and mitochondrial 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, 14-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 stepwise 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 –rRNA Yeast Kits ensure consistently high performance with RNA amounts ranging from as little as 1 ng to 1 μg. QIAseq FastSelect can be used with RNA from fresh samples, as well as degraded RNA, and delivers 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, 14-minute inline step (figure  QIAseq FastSelect Kit workflow). This is dramatically faster than alternative RNA depletion kits, which require pre-treatment protocols involving more than 25 steps and 2 hours to complete.

Simply add QIAseq FastSelect reagent to the RNA sample, perform fragmentation (if required), stepwise cool the reaction from 75°C to 25°C for 14 minutes and then complete the remaining library preparation steps. QIAseq FastSelect works with or without RNA fragmentation, providing the flexibility to use RNA from degraded RNA samples, or high-quality RNA as part of a standard RNA-seq library construction workflow.

See figures

Applications

QIAseq FastSelect delivers rapid, reliable RNA removal from degraded yeast RNA and fresh sample RNA sources. QIAseq FastSelect –rRNA Yeast Kits are available in a variety of different formats and sizes to suit your specific applications.

Supporting data and figures

FAQ

What RNAs has QIAseq FastSelect been designed to remove?

QIAseq FastSelect Yeast

Saccharomyces cerevisiae and Saccharomyces pombe

  • Cytoplasmic (35S [made up of 25S, 18S & 5.8S], 25S, 18S, 5.8S, and 5S)
  • Mitochondrial (21S, 15S, ACI60_gr01 [large subunit ribosomal RNA], and ACI60_gr02 [small subunit ribosomal RNA])

QIAseq FastSelect Plant

Arabidopsis thaliana and Arabidopsis lyrata

  • Cytoplasmic (5.8S, 18S and 25S)
  • Mitochondrial (5S, 18S and 26S)
  • Chloroplast (4.5S, 5S, 16S and 23S)

 

FAQ ID -147910
How does QIAseq FastSelect work?

The QIAseq FastSelect works by prevented cDNA synthesis of unwanted RNAs during library prep.

To enable this, prior to RNA heat fragmentation, the FastSelect reagent is directly combined with total RNA (1 ng – 1 µg) and the library-prep–specific buffers. Heat fragmentation is then performed, and the reaction temperature is gradually cooled to room temperature. Following this, the remaining library-prep–specific steps are followed. There is no need to perform any type of enrichment on the total RNA samples.

FAQ ID -147907
What RNA range has the QIAseq FastSelect been tested with?

QIAseq FastSelect has been tested with 1 ng – 1 µg of total RNA. The amount of total RNA required for a stranded RNA library prep depends on the selected library prep kit.

FAQ ID -147914
I am using an RNA library prep kit that is not listed in the QIAseq FastSelect handbook. Will QIAseq FastSelect work with my kit?

Please contact technical support.

FAQ ID -147913
What library prep kits has QIAseq FastSelect been tested with?

QIAseq FastSelect has been tested with the QIAseq Stranded Total RNA Lib Kit (QIAGEN, cat. no. 180743, 180745), TruSeq® Stranded (Illumina cat. no. 20020594, 20020595), NEBNext® Ultra™ II Directional (New England Biolabs cat. no. E7760S, E7760L) and KAPA® RNA HyperPrep (Kapa Biosystems cat. no. KK8540, KK8541). Generally speaking, QIAseq FastSelect is compatible with any stranded RNA library prep kit that begins with heat fragmentation of RNA. For additional kits, please contact technical support.

FAQ ID -147912
Will QIAseq FastSelect work in other species?

Yes. Based on sequence homology of the rRNA between the species, rRNAs in other species will be removed.

FAQ ID -147911
Can QIAseq FastSelect Plant or QIAseq FastSelect Yeast be combined with FastSelect 5S/16S/23S?

Yes, using the FastSelect 5S/16S/23S protocol, FastSelect Plant, FastSelect Yeast, or any other version can be added.

FAQ ID -147915
Is it possible to test the efficiency of FastSelect rRNA removal by using a Bioanalyzer, etc.?

No, it is not possible to test the efficiency of the FastSelect reaction by running a portion of the eluate from the bead cleanup on a Bioanalyzer®, TapeStation®, Fragment Analyzer®, etc. FastSelect works by inhibiting reverse transcription of rRNA, which doesn’t occur until the first-strand synthesis reaction during library prep.

FAQ ID -147916
Is QIAseq FastSelect truly as easy as combining a reagent with RNA and ramping down the temperature?

Yes, it is truly that easy. In a single 14 min step, QIAseq FastSelect dramatically reduces rRNA. Routinely, 95–99% depletion of rRNA is observed.

FAQ ID -147909