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Product Information
Cellecta’s DriverMap Adaptive Immune Receptor (AIR) RNA Spike-in Controls are a collection of synthetic mRNA constructs that may be used as universal controls for both commercial and home-brewed AIR sequencing assays based on multiplex RT-PCR or 5’-RACE (SMART) PCR techniques. These first-in-kind RNA spike-in controls are designed to ensure consistent and reproducible T-cell Receptor (TCR) and B-cell Receptor (BCR) clonotype profiling. They contain nearly full-length V(D)JC structures that represent all seven TCR (TRB, TRA, TRG, TRD) and BCR (IGH, IGK, and IGL) chains (Figs. 1 and 2).
Product Format
DriverMap AIR RNA Spike-in controls are available in two formats: Premixed Controls or Triplex Isoform Pools for use in TCR- or BCR-NGS-based AIR profiling, according to your specific applications.
Premixed Controls
48 BCR constructs with each of 16 BCR Isoforms containing three CDR3 variants for each isoform (Cat #: DMAIR-HBR-SC-M)
39 TCR constructs with 13 TCR isoforms with three CDR3 variants for each isoform. (Cat #: DMAIR-HTR-SC-M)
The three CD3 variants from each Isoform Pool are mixed in a 16:4:1 ratio present in three discrete concentrations of 4000, 1000 and 250 molecules (i.e., UMIs)/µl.
Triplex Isoform Pools
16 BCR isoform pools (Cat #: DMAIR-HBR-SC-P)
13 TCR isoform pools (Cat #: DMAIR-HTR-SC-P)
Each set has three constructs with the same V(D)JC isoform receptor structure, and each construct has a unique CDR3 sequence. The sequences of the three CD3 variant constructs within each pool differ from each other by 3 unique single-nucleotide mutations, and the three variants are mixed at a 1:1:1 ratio, each with 2,500 molecules/µl.
Applications for DriverMap AIR RNA Spike-in Controls
Ensure Sensitivity and Specificity: Premixed Controls may be spiked into your experimental sample to measure the accuracy and linear range of your PCR assay.
Error Quantification:Premixed Controls may be used to assess sequencing errors introduced at the RT, PCR and NGS stages.
Sample Quality Control:Triplex Isoform Pools or Premixed Controls may be used to assess sample quality such as in degraded FFPE samples.
Cross-Contamination Detection: Triplex Isoform Pools can measure cross-contamination between different samples.
Fig 1: 48 BCR mRNA spike-in constructs for IGH, IGK and IGL chains are shown. Each construct has three variations in the CDR3 region that differ by three nucleotides in a fixed position.
Fig 2:39 TCR constructs, each representing unique TRBs, TRAs, TRGs, and TRDs are shown.Each construct has 3 variations in the CDR3 region that differ by 3 nucleotides in fixed positions.
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