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Journal of Biomolecular Techniques : JBT logoLink to Journal of Biomolecular Techniques : JBT
. 2020 Aug;31(Suppl):S6.

Nanoscale Automation of a Full-Length RNA Sequencing Library Preparation Method on a Microfluidic Circuit

Thomas Goralski 1, Julie Alipaz 1, Joel Brockman 1, Sangpen Chamnongpol 2, Brian Fowler 1, Jennifer Geis 1, Christopher Kubu 1, Raphael Kung 1, Benjamin Lacar 1, Charles Park 1, Michael Phelan 1, Gang Gang Sun 1, David King 1
PMCID: PMC7424891

Abstract

RNA sequencing (RNA-seq) is the gold standard of expression profiling methods. Our Advanta ™ RNA-Seq NGS Library Prep Kit is an elegant microfluidics-based solution that automates many tedious hands-on steps including poly(A) RNA selection. Advanta RNA-Seq generates up to 48 full-length stranded RNA-seq barcoded libraries from as low as 10 ng of total RNA. Nanoliter reaction volumes significantly reduce reagent consumption to lower overall costs per sample. These barcoded libraries can be pooled prior to bead cleanup steps and subsequent PCR amplifications to further decrease reagent consumption and reduce hands-on time. The results from an internal analytical study and a comparison to results obtained from the Illumina® TruSeq® Stranded mRNA Kit are presented in this poster. Our internal analytical study used a total of 917 samples (Universal Human Reference RNA standard and human brain RNA) at 10 and 100 ng of input. All samples generated genome mapping rates of >80%, with rRNA reads of <10%. Technical replicate correlations were observed to be >98% in all conditions. In addition, we found that a comparable number of genes are detected in UHRR and human brain RNA from libraries prepared with the Advanta RNA-Seq NGS Library Prep Kit and the Illumina TruSeq Stranded mRNA Kit, from both 10 ng and 100 ng starting sample inputs. The Advanta RNA-Seq NGS Library Prep Kit provides an automated RNA-seq library prep solution that substantially minimizes manual pipetting steps and hands-on time and increases walkaway time. Nanoliter-scale reaction volumes significantly decrease reagent consumption to reduce the per-sample costs of RNA-seq library construction, providing an attractive value proposition for high-throughput core laboratories. For Research Use Only. Not for use in diagnostic procedures.


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