Overview

The Single Cell Fixed RNA Profiling (FRP) workflow measures RNA levels in samples (single cells or nuclei) fixed with formaldehyde, using probes targeting the whole transcriptome. After whole transcriptome probe panels are added to the sample, each probe pair hybridizes to its target gene and is then ligated. Libraries are generated from the barcoded probes and sequenced. The final sequencing library structure is similar to Gene Expression solutions for fresh frozen tissue, but the synthetic probe DNA is sequenced rather than the cDNA of a captured transcript. Our Core offers this service. Investigators will need to prepare and fix their cells following 10X instructions for this assay.

Technology

We use the following commercially available kits and platforms for sequencing library preparation and sequencing.

• Chromium Fixed RNA kit, Human/Mouse Transcriptome, 4 rxn
• Chromium Next GEM Single Cell Fixed RNA Sample Prep Kit, 16 rxn
• Chromium Next GEM Chip Q Single Cell Kit, 48 rxn
• Dual Index Kit TS Set A, 96 rxn

Key Features

• Gene expression analysis on fixed cells or nuclei
• Process cells fixed with formaldehyde-fixed samples
• Increased sample throughput in a single GEM well with multiplexing

Third-Party Resources

Cell Ranger and Loupe Browser | Single cell data analysis and data visualization tools

The Loupe Browser is a desktop application that provides interactive visualization functionality to analyze data from different 10x Genomics solutions. Cell Ranger and Loupe Browser

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