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DriverMap™ Genome-Wide Targeted Expression Profiling Kit

The development of the DriverMap Targeted RNA Sequencing 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 that–without intermediate steps– provides robust measurements of each expressed gene from total RNA. Even down to single-cell levels of 10 pg, the assay produces a highly quantitative and reproducible NGS readout that has a range of measurement over 5-orders of magnitude.

  • 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

 

As a result of an intensive primer design and testing process that included extensive empirical analysis of multiplex PCR primer mixes, we generated a complete set of validated multiplex RT-PCR primers providing reproducible amplification of all 19,000 human protein-coding transcripts (one cDNA fragment per gene) directly from total RNA. mRNA enrichment is not needed, and, for blood samples, no globin depletion is required. NGS of the amplified products quantifies the relative levels of each transcript using the targeted amplicons as a reference for sequencing deconvolution.

The result is a highly quantitative, targeted RNA expression profiling assay that

  • Accurately measures levels of gene expression with great reproducibility (R-squared values > 0.9)
  • requires only total RNA as input for a multiplex PCR reaction so it is simple and fast (1-day) to run
  • generates defined single-amplicon sequencing results per gene, which makes deconvolution straightforward and easily comparable across samples

DriverMap Assay: Key Features & Benefits

Key FeaturesBenefits
Multiplex RT-PCR-NGS methodNo rRNA or globin depletion required.
Use directly with total RNA.
RNA Input10 pg total RNA (single-cell) minimum
1 ng - 50 ng is optimal
Sensitivity/ Dynamic RangeLinear data up to 10^5- fold dynamic range
10- to 100-fold more sensitive than RNA-Seq or GeneChip methods
SpecificityNo background (sequencing data)
Low cross-amplification of mouse RNAs
Single-tube protocol96 samples per day (2 hours hands-on time)
Result ValidationConventional qRT-PCR using the same DriverMap PCR primers

Cellecta can also run the DriverMap Assay on your samples for you. For more information, please see our DriverMap Service Page.

DriverMap-Brochure

Protocol

Additional material such as scientific posters, video testimonials and other resources are available on the DriverMap Targeted Expression Profiling Service page.