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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)
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KitIndexAccessories
QIAseq UPX 3' Transcriptome Kit
Index
QIAseq UPX Cell Lysis Kit (6 ml)
Samples
96
4 x 96
384
QIAseq UPX 3’ Transcriptome Kitsは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。
✓ オンライン注文による24時間年中無休の自動処理システム
✓ 知識豊富で専門的な製品&テクニカルサポート
✓ 迅速で信頼性の高い（再）注文
特徴
- 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
製品詳細
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。
パフォーマンス
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").
図参照
原理
QIAseq UPX 3' Transcriptome Kits enable Sample to Insight, high-throughput NGS of polyadenylated RNAs from cell lysates or isolated RNA on Illumina NGS instruments. Kits are intended for library construction and analysis of cell pellets (up to 100 cells) or 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. The GeneGlobe-integrated RNA-seq Analysis Portal enables primary mapping and differential expression analysis.
操作手順
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 RNA from cell lysates 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 low-input gene expression analysis.
図参照
アプリケーション
- High-throughput screening applications (e.g., toxicology research)
- Differential gene expression analysis
裏付けデータと数値
Library QC and Quantification.
A portion of the 11 μl sequencing library was used as the starting material for the library QC and quantification. Library QC using an Agilent Bioanalyzer is shown with a peak at 425 bp. Real-time PCR-based methods provide an accurate quantification of complete RNA-seq libraries with full adapter sequences. As a result, QIAGEN’s QIAseq Library Quant Array Kit (cat. no. 333304) or Assay Kit (cat. no. 333314), which contains laboratory-verified forward and reverse primers together with a DNA standard, is highly recommended for accurate quantification of the prepared library. Use 1 nM RNA-seq libraries as input for the denaturation procedure to ultimately load 3 pM for the MiSeq (V3 kit) and 1.2 pM for the NextSeq.
リソース
Supplementary Protocols (2)
MSDS (2)
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)
テンプレートファイル (1)
次世代シークエンシング (1)
Safety Data Sheets (1)
Certificates of Analysis (1)
Template Files (1)
Kit Handbooks (1)
Quick-Start Protocols (3)
Next Generation Sequencing (1)