QIAseq miRNA Library QC PCR Panel and Assays
For evaluating RNA sample quality prior to miRNA/small RNA NGS library preparation and for assessing NGS performance post-sequencing
QIAseq miRNA Library QC PCR Assays and Arrays enable rigorous quality control of both sample quality before NGS library preparation as well as NGS performance post-sequencing. Amplification of the QIAseq miRNA Library QC Spike-ins – a comprehensive set of spike-ins added during RNA isolation or to isolated RNA – allows assessment of the reproducibility and linearity of the NGS reads mapped to these exogenous sequences, ensuring optimal miRNA sequencing results.
QIAseq miRNA Library Kits are recommended for gel-free library construction miRNA, piRNA and small RNA on Illumina and Thermo Fisher Scientific NGS instruments.
QIAseq miRNA Library QC PCR Panel and Assays are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
The QIAseq miRNA Library QC PCR Panel Kit assesses the quality of RNA isolation for small RNA next-generation sequencing (NGS) by providing spike in controls with a qPCR panel that allows researchers to monitor for reproducibility between miRNA isolations, the presence of enzymatic inhibitors and nucleases, sample assessment for hemolysis (important for serum and plasma miRNA identification) and a thorough QC of the NGS data by assessing the reproducibility and linearity of the reads mapped to these exogenous sequences.
The kit is compatible with many samples types but is especially useful for challenging samples with low RNA content such as serum, plasma, urine and other biofluids.
Addition of QIAseq miRNA Library QC Spike-Ins during RNA isolation enables monitoring of the comparability and reproducibility of the whole process from RNA isolation to NGS data analysis.
QIAseq miRNA Library QC Spike-Ins can also be added directly to RNA samples before small RNA library preparation. For more accurate ratios of spike-ins vs. endogenous miRNAs in other sample types or when using isolated RNA samples, experimental titration of QIAseq miRNA Library QC Spike-Ins is recommended.
The protocol consists of 5 steps:
1. Addition of 52 QIAseq miRNA Library QC Spike-Ins to the samples during RNA isolation
2. cDNA synthesis, including UniSp6 Spike-In
3. qPCR reactions
4. Sample evaluation and NGS of accepted samples
5. Evaluation of the QIAseq miRNA Library QC Spike-Ins data
Analysis and interpretation of data
The QIAseq miRNA Library QC Spike-In controls enable you to monitor the technical quality of the RNA isolation and cDNA synthesis and the presence of PCR inhibitors in the sample.
The GeneGlobe data analysis center for QIAseq miRNA library kits (https://www.qiagen.com/us/shop/genes-and-pathways/data-analysis-center-overview-page/) will automatically align and report on the QIAseq miRNA spike-ins in addition to the aligned small/miRNA/piRNA from your sample.
qPCR Panel and Assay interpretation:
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