RNA-Seq (100 NG)
RNA-Seq (Quantification) is used to characterize the gene expression of organisms under various conditions. A challenge in applying RNA-Seq, however, has been the relatively large amount (1 μg total RNA) of required samples. BGI has overcome this obstacle with a technique that reduces the required input RNA to only 100 ng while still delivering high-quality results.
A comparative RNA-Seq analysis using MAQC samples with 1ug and 100 ng input RNA samples demonstrated high reproducibility of the technique, significant correlation between data generated, as well as good concordance with data qPCR. This breakthrough facilitates research dealing with limited samples, such as stem cell studies, paleoarcheology, evolutionary biology and cancer research.
While the technique is still in its research and development stages, we have made it available to pharma and biotech customers for human, mouse, rat, and monkey research.
BGI’s 100 ng RNA-Seq (Quantification) service offers:
- Reduced sample requirement to as low as 100ng total RNA
- Greater reproducibility
- Greater specificity: more accurate, no cross-hybridization, no restrictions to the available probes
- Greater sensitivity: detects more genes, wider dynamic range
- Strong bioinformatics analysis capabilities on RNA-Seq data
- Results compatible with common analysis software
To learn more, download the RNA-Seq with Low Input RNA Amount (100 ng) brochure.
Universal Human Reference RNA (UHRR) and Human Brain Reference RNA (HBRR) were sampled on RNA-Seq by Illumina HiSeqTM 2000 sequencer (50SE). Samples were prepared by following Illumina “mRNA Sequencing Sample Preparation Guide” using 100ng and 1ug of input total RNA. RNA libraries were constructed by two operators (batch 1 and batch 2). Libraries were then pooled in different lanes for sequencing.
The MA plot and the scatter plot were used to evaluate the technical replicates of 100ng RNA-Seq (Quantification). The median value from MA plot was around zero, and the R value of the correlation analysis was 0.96, indicating high reproducibility between technical replicates.
We compared the qPCR data with the data generated by 100ng RNA-Seq to examine the accuracy of the technique. The R value was above 0.9, which indicated that the 100ng RNA-Seq data were in strong concordance with the gold standard (qPCR). This study demonstrated that with BGI’s technique, the amount of input total RNA for RNA-Seq (Quantification) can be significantly reduced without compromising performance.
RNA-Seq (Quantification) based on NGS technology has recently become a new method for characterizing gene expression profiling. It can simultaneously interrogate and precisely measure the expression levels of tens of thousands of samples. When compared to array based methods, RNA-Seq shows greater sensitivity, accuracy and broader dynamic range. Currently, RNA-Seq (Quantification) is widely used in the areas of disease research, biomarker discovery, drug response, pharmacokinetics, pharmocotoxicology, and personalized healthcare.
- Sample condition: RNA samples after DNAase treatment;
- Sample quantity demanded: ≥100ng;
- Sample concentration: ≥ 5 ng/μL;
- Sample purity: OD260/280 =1.8~2.2.