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 want information for delta/gamma TCRs from whole blood, using a 7-chain assay would not be advised since the delta/gamma frequency is naturally lower in whole blood (~5%) for most individuals. For this case, we would either recommend a 1- or 2-chain RepSeq+ 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|