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 then 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. However, when using less than 10 ng of total RNA, the measurements for low abundant transcripts may be less reproducible.
Main advantages of DriverMap™ EXP hGW19K Assay:
- 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.
Related Products and Services:
- DriverMap™ AIR TCR/BCR Profiling Kit (human/mouse RNA/DNA) is designed to generate comprehensive AIR CDR3 repertoire profiling of all seven TCR (TRA, TRB, TRD, and TRG) and/or BCR (IGH, IGK, and IGL) chains from RNA or DNA samples. AIR RNA assay provides higher sensitivity in detecting full-length (CDR1-CDR2-CDR3) or CDR3 clonotypes than AIR DNA assay, especially from small biological samples. Both AIR RNA and AIR DNA assay could be effectively used together to identify antigen-induced clonotypes. For more details, please refer to the AIR Technical Guide.
- DriverMap™ EXP Human Genome-wide 19K Dried Blood Microsamples Profiling Kit allows profiling from 30 µl of dried blood collected by Mitra® devices with VAMS® technology. Expression profiling in dried blood is a convenient strategy for longitudinal analysis of blood samples which could be collected directly by patients without going to a blood collection facility.
Cellecta also provides a DriverMap™ Expression Profiling Service. For more information, please visit our website:
DriverMap EXP Service Webpage
Please read the entire user manual before proceeding with your experiment.
PLEASE NOTE: The purchase of all Cellecta Products is covered by Cellecta’s standard Terms and Conditions of Sale as described on Cellecta’s website, and selected Products containing particular technology or having certain features are also subject to restrictions on use as outlined in the Label License section of our website. Please review these Terms and Label License Restrictions before opening and using your Product and, if you are not able to abide by the restriction, contact Cellecta to return the item to Cellecta for a full refund.
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