Gordon Research Conference 2018: Liquid Biopsy for Cancer
SMART ligation-free tools for sequencing coding and noncoding RNA from liquid biopsy samples
Small and long noncoding RNAs play a major role in the regulation of gene expression and disease. Obtaining an accurate portrait of their expression levels from small sample inputs carries potential for both the fulfillment of basic research objectives and the development of novel therapeutics and clinical diagnostic solutions. Towards this end, we have built upon the sensitivity of our SMART technology to develop the SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (Pico kit). The Pico kit relies on random priming to synthesize cDNA from both polyadenylated and non-polyadenylated RNA, thus ensuring a full RNA representation from even the most challenging samples, such as highly degraded RNA from FFPE or cfRNA. In addition, we developed a novel technology, ZapR, which allows for the removal of ribosomal RNA (rRNA)-derived cDNA after reverse transcription and library amplification. The Pico kit enables strand-specific transcriptome analysis from very low amounts of total RNA or cells, but it does not allow for the analysis of small RNAs, such as miRNAs, piRNAs, etc. To address this limitation, we developed the SMARTer smRNA-Seq Kit for Illumina, a ligation-free approach for the preparation of small RNA sequencing libraries that leverages 3′ RNA polyadenylation followed by cDNA synthesis and template switching. This approach minimizes sample representation bias and is sensitive enough to accommodate inputs of as little as 1 ng of total RNA. The combination of the SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian and the SMARTer smRNA-Seq Kit for Illumina provide a complete toolkit for accurate, sensitive, and reproducible detection of coding and noncoding RNAs of any size from the most challenging samples.
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