For rapid 5S/16S/23S rRNA removal for RNA-seq library preparation from bacterial RNA samples

Features

Pan-bacterial 5S/16S/23S rRNA removal from fragmented or full-length RNA for metatranscriptomics studies

Novel RNA removal mechanism that doesn't involve hybrid capture or enzymatic digestion

Combine with QIAseq FastSelect –rRNA HMR for ribosomal RNA removal from human, mouse and rat samples for host-microbe studies

Compatible with QIAGEN, Illumina, NEB and KAPA stranded RNA-seq library preparation kits

In-silico design predicts blocking of >95% of all 5S, 16S and 23S rRNA database sequences

Product Details

QIAseq FastSelect –5S/16S/23S Kits use a novel method to remove highly abundant bacterial 5S/16S/23S rRNA from your RNA-seq library. QIAseq FastSelect –5S/16S/23S was specifically designed for complex community microbial samples from soil, water, stool and sludge, which have numerous and diverse bacterial populations. The rRNA coverage provided by this kit is based on SILVA 16S sequences (nearly 600,000 entries), SILVA 23S sequences (nearly 170,000 entries) and 5S rRNA database sequences (over 7200 entries), and in-silico modeling shows coverage of >95% of all known 5S, 16S and 23S rRNA sequences.

QIAseq FastSelect –5S/16S/23S has been tested with QIAGEN, Illumina and NEB RNA-seq library kits and is compatible with all major methods of RNA-seq library preparation. The simple 14-minute protocol and bead cleanup integrates seamlessly with all common RNA-seq library kits, and can be used on fragmented or full-length (non-fragmented) RNA.

For removal of human, mouse, rat and other mammalian rRNA, please see our QIAseq FastSelect –rRNA HMR Kits.

Also, check out our QIAseq Stranded RNA Library Kits for robust RNA-seq library preparation.

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 this solution for the first time? Request a trial kit to evaluate.

Performance

In accordance with QIAGEN’s ISO-certified Quality Management System, each lot of the QIAseq FastSelect –5S/16S/23S Kit is tested against predetermined specifications to ensure consistent product quality.

QIAseq FastSelect –5S/16S/23S Kits provide robust rRNA removal from a variety of sample types.

QIAseq FastSelect –5S/16S/23S Kits deliver highly efficient and reproducible rRNA removal.

QIAseq FastSelect –5S/16S/23S Kits demonstrated highly efficient removal of rRNA and superior performance compared to alternative methods.

QIAseq FastSelect –5S/16S/23S Kits are compatible with a wide range of RNA-seq library kits.

Principle

Used after bacterial RNA extraction, QIAseq FastSelect –5S/16S/23S is a pan-bacterial rRNA removal kit designed to remove 5S, 16S and 23S rRNA from complex bacterial community samples. Our comprehensive rRNA removal reagent has been designed using 16S (nearly 600,000 entries), 23S (nearly 170,000 entries) and 5S rRNA sequences (over 7200 entries). In-silico modeling predicts >95% rRNA removal of all known 5S, 16S and 23S rRNA sequences.

QIAseq FastSelect –5S/16S/23S can accommodate RNA amounts ranging from as little as 20 ng to 1 µg, with consistently high performance, and is compatible with low-quality, highly fragmented or high-quality full-length RNA. Since QIAseq FastSelect –5S/16S/23S does not use enzymatic digestion or hybrid-capture procedures, the fast, simple workflow results 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-treatments involving hybrid-capture or enzymatic removal of unwanted RNA. Our unique QIAseq FastSelect procedure is significantly faster and is compatible with most RNA-seq library kits. QIAGEN FastSelect –5S/16S/23S has been tested with QIAGEN, Illumina and NEB stranded RNA-seq library kits.

QIAseq FastSelect –5S/16S/23S seamlessly integrates with your existing RNA-seq library preparation, providing RNA removal in a single, 14-minute step, which is followed by bead cleanup. Prior to RNA heat fragmentation (which is optional and dependent upon the library preparation kit and sample type), QIAseq FastSelect –5S/16S/23S removal reagent is directly combined with total RNA and fragmentation buffer. After optional fragmentation, the reaction temperature is stepwise cooled from 75°C to 25°C over 14 minutes, followed by a bead cleanup. The RNA is then ready for reverse transcription. This is dramatically faster than other RNA depletion methods, which require more than 25 steps and approximately 2 hours to complete.

Applications

QIAseq FastSelect –5S/16S/23S delivers rapid, reliable rRNA removal for bacterial isolates, as well as complex bacterial communities.