High-precision analysis for confident results and deep insights
iRepertoire’s RepSeq+ service utilizes our dam-PCR technology, which allows for any combination of TCR and BCR chains (TCR-alpha, TCR-beta, TCR-delta, TCR-gamma, BCR-IgHeavy, and BCR-kappa/lambda) to be amplified within a single reaction.
Multichain analysis in one single reaction mix is beneficial in that it saves rare and/or precious samples by using less RNA than traditional methods. Dam-PCR also allows for the detection of unique mRNA transcripts, which provides reduced bias, better error removal, and more precise data on clonotype frequency.
One chain versus multi-chain
With RepSeq+, the sequencing reads devoted to a particular chain are determined by the natural state of the sample, which allows for calculations of BCR to TCR expression and clonotype ratios; this should be taken into consideration prior to chain selection for examination in the assay. For instance, if you would like to know the frequency of expressed delta and gamma RNAs in the context of the rest of the repertoire, a RepSeq+ 7-chain assay is the optimal way to discover this.
Conversely, if you want in-depth information about delta and gamma clonotype diversity, using a 7-chain assay would not be advised as the delta/gamma frequency is naturally lower in whole blood (~5% on average). For this case, we would either recommend a 1- or 2-chain RepSeq+ analysis or analysis, or the use of our more inclusive, single chain RepSeq service, which relies on iRepertoire™’s arm-PCR technology.
RNA sample submission requirements
RepSeq+ | Dam-PCR
|Minimum concentration||Minimum mass||Optimum concentration||Optimum mass||Minimum volume|
|FFPE||5 ng/uL||75 ng||20 ng/uL||300 ng||15 uL|
|Solid Tissue||5 ng/uL||75 ng||50 ng/uL||1000 ng||15 uL|
|PBMC / Whole Blood||5 ng/uL||75 ng||50 ng/uL||1000 ng||15 uL|
|Sorted Cell Populations||1 ng/uL||15 ng||10 ng/uL||200 ng||15 uL|