QIAseq UPX 3’ Transcriptome Kits

微量の RNA からハイスループット 3’トランスクリプトーム解析を実現

Features

  • 1~100 個の細胞または 10 pg~1 ng の精製 RNA から対応
  • LNA 技術を採用して、精度、特異度、感度を向上
  • 分子バーコード(UMI)により増幅バイアスが除去され、正確な遺伝子発現を実現
  • セル ID とサンプルインデックス(プレート ID)の組合せで、最大 18,432 サンプルの転写産物を同時にシークエンス解析可能
  • GeneGlobe によるクラウドベースの無償データ解析ツールを提供
QIAseq UPX 3' Transcriptome Kit (96)

Cat. No. / ID: 333088

For 3' gene expression analysis from cells or mRNA of 8–96 samples at one time; each kit contains cell lysis solution, buffers, oligos and enzymes lyophilized in a 96-well plate which can be easily separated for processing 8–96 samples in parallel (this format is ideal for both small and large sample sizes)
¥307,000
KitIndexProduct
QIAseq UPX 3' Transcriptome Kit
Index
QIAseq UPX Cell Lysis Kit (6 ml)
Size
QIAseq UPX 3' Transcriptome Kit (96)
QIAseq UPX 3' Transcriptome Kit (96-M)
QIAseq UPX 3' Transcriptome Kit (384)
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QIAseq UPX 3’ Transcriptome Kits は分子生物学実験用です。疾病の診断、治療または予防の目的には使用することはできません。

Product Details

QIAseq UPX 3' Transcriptome Kits により、シングルセルレベルの微量 RNA (1-100細胞 / 細胞ペレット、1 pg~10 ng 精製RNA)から、3' RNA シークエンスによるハイスループットな遺伝子発現解析が可能です(イルミナシークエンサー対応)。
本キットは、96 ウェルプレート(切り離し可能)、もしくは 384 ウェルプレートで構成されています。それぞれプレートの底には凍結乾燥した逆転写プライマーが添加済みで、使いやすさも考慮されています。1ウェルにつき 1~100 個の細胞、もしくは 10 pg~1 ng の精製 RNA を加え、ウェル内で逆転写反応と同時にセル ID および分子バーコード(UMI)付加を行います。cDNA 合成後、各プレートを1本のチューブにまとめ、ライブラリー調製をすることが可能です。必要な試薬は全てキットに含まれております。ご希望の index キットと組合せてご使用ください。

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 (see figures " Streamlined and Reproducible Workflow Starting from 10 pg to 1 ng of Total RNA", " Consistency of Detection with QIAseq UPX 3’ Transcriptome Kits and " Library QC and Quantification").
See figures

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 (see figures " Innovative and Optimized QIAseq UPX 3' Transcriptome Kit Workflow" and " QIAGEN’s Sample to Insight QIAseq UPX 3' Transcriptome Workflow").

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.
See figures

Applications

  • High-throughput screening applications (e.g., toxicology research)
  • Single-cell analysis
  • Differential gene expression analysis

Supporting data and figures

Resources

キットハンドブック (1)
クイックスタートプロトコール (3)
Part 1: Cell lysis and reverse transcription
Part 3: Adapter ligation and universal PCR
Part 2: Template amplification and fragmentation, end-repair, and A-addition
テンプレートファイル (1)
次世代シークエンシング (1)