For high-throughput 3' transcriptome analysis from up to 10 ng of purified RNA, cell lysates and single cells using next-generation sequencing

Features

Start with 10 pg to 10 ng of isolated total RNA, cell lysates or single cells

UPX tagging allows 4608–18,432 samples per single sequencing lane

UMIs eliminate library amplification bias for accurate gene expression

LNA-enhanced chemistry for increased accuracy, specificity and sensitivity

Includes access to the RNA-seq Analysis Portal for human, mouse and rat samples

Product Details

QIAseq UPX 3' Transcriptome Kits enable high-throughput gene expression analysis from single cells and isolated total RNA. Each kit is enough for 96 or 384 libraries with kit formats comprised of either 96-well multi-break plates or 384-well single-use plates with reverse transcription primers already lyophilized in the bottom of the plates for ease of use. To build libraries for RNA-seq, simply add 1 to 100 cells into each well or up to 10 ng of total RNA to reconstitute a reverse transcription reaction. After cDNA synthesis, combine each plate into a single tube to complete the library preparation. All reagents necessary for starting with cells (such as lysis reagents) or RNA are included in the kit.

Analyze 3’ RNA-seq data with ease using the GeneGlobe-integrated RNA-seq Analysis Portal – an intuitive, web-based data analysis solution created for biologists and included with QIAseq UPX 3’ Transcriptome Kits.

Performance

The QIAseq UPX 3' Transcriptome Kit presents a unique advantage where each cell is tagged with a unique ID (up to 384 different IDs) and each RNA molecule is tagged with a Unique Molecular Index (UMI) during reverse transcription. Following reverse transcription with integrated template switching, all individually tagged cDNAs can be combined, which enables all subsequent library construction steps to be performed in a single tube – saving significant time and library prep costs. QIAseq UPX 3’ Transcriptome Kits provide a convenient, streamlined and reproducible library construction workflow starting with as little as 10 pg total RNA.

Principle

QIAseq UPX 3' Transcriptome Kits enable Sample to Insight, high-throughput NGS of polyadenylated RNAs from single cells on Illumina NGS instruments. Kits are intended for library construction and analysis of cell pellets (up to 100 cells) and purified RNA (10 pg to 10 ng). QIAseq UPX 3' Transcriptome Kits present an innovative advantage in that during reverse transcription, each cell is tagged with a unique ID (up to 384 different IDs) and each RNA molecule is tagged with a Unique Molecular Index (UMI). Following reverse transcription with integrated template switching, all individually tagged cDNAs can be combined, which enables all subsequent library construction steps to be performed in a single tube. This prevents sample mixup, saves substantial time and dramatically reduces library prep costs. During subsequent amplification and library construction up to 48 different sample IDs can be assigned. Together, the combination of cell IDs and sample IDs enables up to 18,432 libraries to be sequenced together. QIAseq UPX data analysis enables primary mapping, single-cell clustering analysis and differential expression analysis.

Procedure

Innovative, optimized workflow

The QIAseq UPX 3' Transcriptome Kit defines a new generation of high-throughput NGS technologies in QIAGEN’s Sample to Insight workflow. Purified RNA or single cells are first reverse transcribed and each RNA molecule is given a Unique Molecular Index (UMI) and well-specific IDs are assigned (up to 384 wells; cell IDs). Following reverse transcription with integrated template switching, all cDNAs are combined, enabling simplified library construction steps to be performed in a single tube. During subsequent amplification and library construction, up to 48 different library indices (sample IDs) can be assigned. The combination of cell IDs and sample IDs enables up to 18,432 libraries to be sequenced simultaneously .

Convenient GeneGlobe data analysis

QIAseq UPX cloud-based data analysis is available via the GeneGlobe Data Analysis Center and provides read alignments, UMI and sample de-multiplexing, with final single-cell or low-input gene expression analysis.

Applications

High-throughput screening applications (e.g., toxicology research)

Single-cell analysis

Differential gene expression analysis