DriverMap™ EXP Human Genome-wide 19K Profiling Kit, V3

Product Description

The DriverMap™ EXP Human Genome-wide 19K Profiling Kit (hGW19K) measures the expression level of almost 19,000 human protein-coding genes by combining highly multiplexed RT-PCR amplification with the depth and precision of Next-Generation Sequencing (NGS) quantitation. It is an easy-to-run, single-tube, single-day assay that can be run directly from total RNA isolated from cells, tissue, or blood, or directly from cellular extract using the DirectCell™ Protocol.

The development of the DriverMap™ EXP hGW19K Assay involved extensive optimization and experimental validation of tens of thousands of primer sets to identify a pool of primers that could be combined in a single multiplex RT-PCR reaction to amplify representative transcript sequences (i.e., conserved transcript regions common to different mRNA isoforms from the same gene) from each of ~19,000 protein-coding genes in the human genome.

The amplified sequences are quantified using NGS on the Illumina platform. The DriverMap™ EXP hGW19K Assay provides optimal quantitative expression data within a 10⁴ – 10⁵-fold dynamic range from 10-50 ng of total RNA and can be used effectively with as little as 1 ng of RNA. 

• An easy-to-run, one-tube, single-day assay allows profiling of transcripts from total RNA isolated from whole blood, PBMC, tissue, or directly from a small number (e.g., 100-50,000) of purified cells without prior RNA isolation
• The assay provides robust, quantitative, and reproducible measurements of the copy number of mRNAs over as many as 4-orders of magnitude differences in expression level
• The assay provides increased sensitivity from a small amount of RNA (1-50 ng) using a set of highly experimentally-validated primers
• The DriverMap EXP protocol employs several mechanisms to minimize amplification of off-target and primer dimer background products
• No mRNA enrichment, rRNA, mitochondrial, beta-globin depletion, or other total RNA processing is required
• The primers are designed using Illumina’s DNA/RNA dual UD Indexes labeling strategy to minimize NGS index-swapping background issues, and amplified products are compatible with 100-150-cycle Paired-End NGS sequencing using Illumina’s NextSeq/NovaSeq instruments
• NGS data can be analyzed using the Salmon software package available online. Cellecta also provides custom data analysis services on request. Please contact us

Gene List

Gene List DriverMap™ EXP Human Genome-wide 19K Profiling Kit, V3

How It Works

Gene-specific primers (GSPs) initiate first strand synthesis. As a result, gene-specific first-strand cDNA templates are synthesized in the RT reaction. The RNA is removed after the RT reaction, and as a consequence, the reaction has much less background.