Fully Automated Sample Preparation Methods for High Throughput Transcriptome Analysis by Arrays and NGS

摘要

NuGEN Technologies is a provider of innovative sample preparation solutions. As such, we offer a wide palette of systems optimized for a variety of sample types such as whole blood RNA, small quantity RNA from different sources as well as highly degraded RNA i.e. from FFPE specimens. The SPIA® (Single Primer Isothermal Amplification) technology allows rapid linear amplification of input RNA even at picogram level and all of our amplification assays generate microgram amounts of cDNA, known as SPIA product which is suitable for analysis i.e. by qPCR. Reagent modules to further prepare the SPIA product for subsequent analyses on microarrays or massive parallel sequencing complement NuGEN's current offering. NuGEN targets are compatible with all leading microarray formats and NGS platforms. Automating a workflow not only helps maximizing sample throughput but also minimizes variance introduced during the assay. We describe fully automated assays on leading liquid handling systems that enable high-throughput target preparation of up to 96 samples using NuGEN systems for different sample types. The NuGEN assays are scripted in a modular form allowing highest flexibility with regard to both, the number of samples processed at a time as well as integration into the laboratory workflow. Starting from total RNA, the entire process through to target preparation for further analysis on microarrays or sequencing platforms can be completed within two convenient workdays. Basic QC-metrics with these fully automated workflows show equal or superior performance over manual handling. These methods are well suited for adoption by core facilities to increase efficiency, reduce operator variation, and improve reproducibility and data quality. Their compatibility with a range of microarrays and sequencing platforms further enhance the flexibility for the core labs that can adopt a single workflow for a range of instruments and applications.