Area of Interest Genomics Analysis NGS Library Prep RNA-Seq RNA-Seq with Microfluidics

Advanta RNA-Seq XT NGS Library Prep Kits 

Transform your RNA sequencing productivity with nanoscale automation

RNA sequencing (RNA-seq) is the standard for hypothesis-free profiling of the transcriptome and is an essential tool for molecular biology laboratories. Designed to drive significant improvement in the RNA-seq workflow, the Advanta™ RNA-Seq NGS Library Prep Kit together with the Juno™ system provides an integrated solution for automated, cost-efficient NGS library prep. 

Blue icon illustrating RNA sequencing application

Fluidigm has developed an elegant microfluidics-based platform for automated RNA-seq library preparation. Leveraging Juno™ and the 48.Atlas™ integrated fluidic circuit (IFC), the workflow supports the simultaneous processing of up to 48 total RNA samples with a one-click script on the instrument. With the launch of the Advanta™ RNA-Seq XT Library Prep Kits we have updated the products to include unique dual index (UDI) adapters, ideal for use on NovaSeq™ platforms, as well as simplified the reagent usage for additional savings on hands-on time and labor costs. 

The Advanta RNA-Seq XT Kits contain sufficient 48.Atlas IFCs (4 per kit), reagents, Control Line Fluid and UDI adapters to generate libraries from 192 total RNA samples, and include either 4 sets of 1–48 UDI barcodes (PN 101-9950) or 2 sets of 1–48 UDI barcodes and 2 sets of 49–96 UDI barcodes (PN 101-9959). 

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The Advanta RNA-Seq XT NGS Library Prep Kits and Juno system advantage 

Advanta RNA SEQ flyer inforgraph

Figure 1. Advanta RNA-Seq NGS XT Library Prep Kit workflow on Juno. Samples and Advanta RNA-Seq XT reagents are added to the 48.Atlas IFC, which is subsequently processed on the Juno instrument. 

The system solution automates many tedious, hands-on steps to generate full-length, stranded libraries from random priming of polyadenylated [poly(A)] RNA present in total RNA samples. The Advanta RNA-Seq XT Kit and Juno workflow improve upon alternative RNA-seq library prep workflows by providing true walkaway automation while reducing reagent, consumables, labor and instrument costs. 

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WORKFLOW 

Advanta RNA-Seq XT workflow, 48.Atlas IFC and Juno enable significant efficiencies 

The Advanta RNA-Seq XT Kits include library preparation reagents, UDI adapters, and the 48.Atlas IFC, a microfluidic device approximately the same size and shape as a standard 96-well plate. Sample and reagent mixes are dispensed into the IFC, which is then placed in the Juno system for automated processing of most of the workflow steps. Because processing on Juno requires no manual intervention and no pipetting, the solution greatly reduces hands-on time as well as consumables cost and plastic waste. 

Figure 2. The 48.Atlas IFC architecture automates multiple workflow steps otherwise performed manually, including poly(A) RNA capture, RNA fragmentation, reverse-transcription, sample-barcode PCR and multiple wash steps. 

WORKFLOW COMPARISON

Example comparing hands-on time and number of manual processing steps for several RNA-seq workflows.

Figure 3. Example comparing hands-on time and number of manual processing steps for several RNA-seq workflows. Comparisons were made between the Advanta RNA-Seq XT NGS Library Prep Kit, TruSeq® Stranded mRNA LP, NEBNext® Ultra II Directional RNA LP and KAPA mRNA HyperPrep LP. All workflows include quantification of 48 samples by qPCR and pooling to obtain the final library for sequencing. The Advanta RNA-Seq XT workflow using the 48.Atlas IFC with Juno system automation enables substantial hands-on time savings relative to the alternative library prep workflows. 

PERFORMANCE

Summary of performance results generated from analytical validation testing. 

Summary of performance results generated from analytical validation testing.

Table 1. Performance characteristics of the Advanta RNA-Seq XT Kit (PN 101-9959) on Juno were assessed in an analytical validation study. Total RNA samples were Universal Human Reference RNA (UHRR; Agilent®) and human adult brain RNA (Brain; BioChain®). The results demonstrate high mapping statistics with a low-percentage reads mapping to rRNA sequences for both RNA types and input amounts of either 10 ng or 100 ng. 

Learn more. Download the Advanta RNA-Seq XT NGS Library Prep Kit Application Note. 

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Unless explicitly and expressly stated otherwise, all Fluidigm products are provided for Research Use Only, not for use in diagnostic procedures. For more information see www.fluidigm.com/legal/notices