DriverMap™ Genome-Wide Targeted Expression Profiling Kit
The DriverMap™ Human Genome-Wide Targeted Expression Profiling Assay enables researchers to simultaneously measure the expression level of almost 19,000 human protein-coding genes in a single assay. By combining highly multiplexed RT-PCR amplification with the depth and precision of next-generation sequencing (NGS) quantitation, the Driver-Map assay provides convenient, comprehensive, highly sensitive, and quantitative measurement of gene expression from total RNA.
Don’t miss out on critical changes in gene expression. Capture the complete gene expression picture of your with Driver-Map targeted expression profiling kit.
- Easy-to-run, one-tube, single-day expression profiling directly from total RNA from any tissue, cell, or blood samples
- Efficient multiplex RT-PCR amplification for comprehensive profiles with greater detection of low abundant transcripts
- Robust, reproducible data from low-input RNA samples—start with as little as 10 pg total RNA
Sensitive, Robust, and Broadly Dynamic, One-Tube Expression Profiling
The development of the Driver-Map 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 PCR reaction to amplify representative transcript sequences from each of 19,000 protein-coding genes in the human genome. The result is a rapid and convenient, single-tube, RT-PCR combined with NGS protocol (Fig. 1) that–without intermediate steps– provides robust measurements of each expressed gene from total RNA—even down to single-cell levels of 10 pg (Fig. 2)—with a highly quantitative and reproducible NGS readout generating up to 5-orders of magnitude digital expression level measurement.
Driver-Map assay has important benefits over other methods
- Functionally validated, GCA-rich PCR primers minimize primer dimerization and cross-reactivity while maximizing specificity and efficacy
- Wider dynamic range and 10- to 100-fold greater sensitivity than microarray and RNA-seq expression analysis methods to detect more low- and medium-abundant transcripts (Fig. 3)
- Spike-in External RNA Controls Consortium (ERCC) calibration standards are included for data QC and normalization
Use Driver-Map Assay Kit with Total RNA from Any Source
Unlike most expression analysis approaches, the Driver-Map assay starts with total RNA. No mRNA enrichment is needed. Any standard RNA sample for RT-PCR is suitable for Driver-Map analysis, e.g. total RNA from cells, frozen tissue, fine needle aspirate (FNA), whole blood, peripheral blood mononuclear cells (PBMCs), and mouse patient-derived xenograft (PDX) isolates. The targeted, validated primers used in the Driver-Map assay specifically amplify the target transcript amplicon sequences with minimal background from other non-target RNAs. As a result, total RNA from blood can be assayed directly without removal of rRNA or globin components. In addition, with the Driver-Map assay, human transcripts can be specifically amplified against the background of mouse RNAs in PDX samples.
The obvious choice for a wide range of applications
- Biomarker Discovery from Whole Blood Samples—use whole blood RNA (e.g., PAXgene) without previous globin and ribosomal RNA depletion.
- Circulating Tumor Cell (CTC) Detection in Whole Blood—no enrichment step needed.
- Tumor Cellular Composition Analysis—comprehensive expression profiles to assess tumor/stromal cell content and profile infiltrating immune cell types
- Patient-Derived Xenograft (PDX) Models— obtain human-specific expression profiles without separation of human cells from model-animal background cells
- Expression Analysis of Cell Lines—easy 96- or 384-well single-tube protocol facilitates identification of the mechanisms of chemical and genetic perturbations
- Expression Profiling of Rare and Limited Cell Populations—comprehensive analysis with limited numbers of cells isolated by FACS, magnetic beads or microfluidics, fine needle biopsies, and laser capture microdissection.
Driver-Map Assay: Key Features & Benefits
|Multiplex RT-PCR-NGS method||No rRNA or globin depletion required.
Use directly with total RNA.
|RNA Input||10 pg total RNA (single-cell) minimum
1 ng - 50 ng is optimal
|Sensitivity/ Dynamic Range||Linear data up to 10^5- fold dynamic range
10- to 100-fold more sensitive than RNA-Seq or GeneChip methods
|Specificity||No background (sequencing data)
Low cross-amplification of mouse RNAs
|Single-tube protocol||96 samples per day (2 hours hands-on time)|
|Result Validation||Conventional qRT-PCR using the same Driver-Map PCR primers|
Cellecta can also run the Driver-Map Assay on your samples for you. For more information, please see our Driver-Map Service Page.