For ultraplex (UPX) gene expression analysis using 3' RNA-seq from single cells and low-input samples

Features

Target up to 1000 genes using a cost-effective, time-saving single-tube library prep

LNA-enhanced chemistry for increased accuracy, specificity and sensitivity

UMIs eliminate library amplification bias for accurate gene expression

Cell tagging and sample indexing enables simultaneous sequencing of up to 147,456 targeted libraries

Includes cloud-based read alignment and single-cell or low-input analysis

Product Details

QIAseq UPX 3' Targeted RNA Panels use unique molecular index (UMIs) and ultraplex (UPX) technology to enable accurate gene expression using 3' RNA-seq from single cells (1–1000), cell pellets and previously isolated total RNA (1 pg to 10 ng). Use our custom builder to design your custom panel for up to 1000 genes. UPX technology allows up to 384 plates of 384 samples (147,456 total samples) to be multiplexed together at one time. QIAseq UPX 3' Targeted Panels are available for 96 or 384 samples. For processing less than 96 samples at a time, use QIAseq UPX 3' Targeted RNA Panel (96) (cat. no. 333041), which is supplied with a breakable plate enabling processing of as few as 24 samples at one time for sequencing. The QIAseq UPX 3' Targeted RNA Panel (96-M) (cat. no. 333042) is for processing 96 samples in parallel 4 times (384 samples). The QIAseq UPX 3' Targeted RNA Panel (384) (cat. no. 333043) is for processing 384 samples simultaneously. QIAseq UPX 3' Targeted RNA Panels are only compatible with QIAseq UPX Targeted RNA Indices, which must be used during library construction for correct indexing.

Performance

Strong reproducibility at ultra-low input RNA amounts

Due to its LNA-enhanced chemistry, the QIAseq UPX 3’ Targeted RNA Panel workflow exhibits strong reproducibility at ultra-low input RNA amounts.

Strong linearity from 10 pg to 1 ng

The QIAseq UPX 3’ Targeted RNA Panel workflow shows strong linearity from 10 pg to 1 ng.

3' RNA-seq from single cells

QIAseq UPX 3' Targeted RNA Panels enable accurate gene expression using 3' RNA-seq from single cells . The kit also enables sucessful identification of cell subtype clusters.

Principle

QIAseq UPX 3' Targeted RNA Panels enable high-throughput next-generation sequencing (NGS) of up to 1000 polyadenylated RNAs on Illumina sequencers. These panels are intended for library construction and analysis of single cells, cell pellets and ultralow amounts of purified RNA. Single cells or isolated RNA are 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, all cDNAs are combined, enabling simplified library construction steps to be performed in a single tube .

Procedure

Innovative, optimized workflow

QIAseq UPX 3' Targeted RNA Panels enable Sample to Insight, high-throughput next-generation sequencing (NGS) of polyadenylated RNAs from single cells on Illumina NGS instruments. In addition, QIAseq UPX is intended for library construction and analysis of cell pellets (up to 100 cells) and purified RNA (10 pg to 1 ng). QIAseq UPX presents 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 index. Following reverse transcription, all cDNAs are 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. Target enrichment technology further enhances the power of QIAseq UPX by enabling users to sequence specific genes of interest, which effectively increases sequencing depth and sample throughput, while minimizing cost. Universal library amplification then assigns up to 384 different sample IDs. Together, the combination of Cell IDs and the Sample IDs enables up to 147,456 targeted libraries to be sequenced together. Ultimately, QIAseq UPX data analysis enables primary mapping, single-cell clustering analysis and differential expression analysis. Collectively, the Sample to Insight workflow of QIAseq UPX defines a new generation of high-throughput NGS technologies.

Applications

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

Single-cell analysis

Differential gene expression analysis