RNA Sequencing (RNA-Seq) is a molecular biology technique that uses next generation sequencing (NGS) to examine the sequence and quantity of RNA in a sample. RNA-Seq profiles transcriptome which is the sum total of all RNA, in order to determine the gene expression pattern of an organism or cell. It quantitively analyzes when and where a gene is turned on/ off, alternative splicing events, and post-transcriptional modifications. This technique is especially useful for garnering information about function of genes, for understanding diseases, pharmacogenomic studies, and personalized medicine.
In the case of iPSCs, an enormously useful tool for research and in future for regenerative medicine there is still considerable heterogeneity associated between iPSC lines due to upstream reprogramming from somatic tissues and downstream directed-differentiation into tissue lineages. As well, a lack of gene expression roadmap for differentiation to various lineages currently limits use of iPSCs in clinical applications. Applied StemCell offers NGS-based, Single-cell RNA-seq services for characterizing pluripotency markers for your iPSCs and differentiated cell lines.
ASC’s Single-cell RNA-seq service for iPSCs, primary cells, cancer cell lines and CRISPR genome edited cell lines includes high-resolution transcriptome screening of a single cell which enables analysis of gene regulation after reprogramming into pluripotent cells as well as lineage-specific gene expression pattern during differentiation. We use:
- Cutting-edge technology for single-cell RNA-seq
- Advanced amplification methods: for efficient amplification, reduced-cost
- Benchmark quality scores and base calling accuracy (Q30) and guarantee (≥ 80%)
Comprehensive processing of samples: amplification, library construction, sequencing, and bioinformatics analysis
- Amplification: 7 working days from verification of sample quality
- Library preparation and sequencing: within 15 working days
- The turnaround for data analysis is project dependent
- Sorted single cells should be stored in one of the two ways (1) in lysis buffer with RNase inhibitor from SMARTer kit (Clontech) or (2) in 1X PBS buffer (excluding calcium and magnesium) containing RNase inhibitor in a total volume of ≤ 2 μl. The stored cell should be frozen in liquid nitrogen and shipped out with dry ice.