For seamless precision in immune profiling. The sCircle® Single Cell Full Length Immunoreceptor Library Kit technology enables full length sequencing of the V(D)J region at single cell level with short-read sequencing.
For full-length immuno receptor sequencing, the cDNA is circularized after standard single cell partitioning, mRNA capturing, barcoding and reverse transcription. Circularization of the cDNA brings the V(D)J region closer to the sequencing primer. The enriched and fragmented cDNA is then used to construct a full-length V(D)J library, which is subsequently analyzed using short-read sequencing.
High-throughput single cell sequencing reveals clonotype frequencies, the full length of the BCR or TCR transcript, their TRA and TRB or BCR IGH/IGL pairing map, as well as the amino acid sequence of their variable regions (CDR3). Here, the most frequent pairing between IGMT V and J genes from TCR are shown.
sCircle® Single Cell Full Length Immunoreceptor Library Kit has significant advantages compared to other methods due to the circularization process of the immunoreceptor cDNA specifically designed to reverse the sequence of the transcript and, through this, allows the full-length of the immune receptor to be sequenced. This results in reads mapped to the full-length sequence of the immunoreceptor gene (BCR or TCR), including the V(D)J region and part of the constant region. It also shows a high immunoreceptor pairing rate, and a high number of immune receptor transcript-specific UMIs detected per cell, rendering both high specificity and sensitivity.
with the HD configuration
Full instruments-independent workflow
PythoN: tissue dissociation NEO: single cell processing
The starting material for sCircle can only be a cell suspension. Users working with tissue may want to buy tissue dissociation reagents e.g. sCelLiVE separately.
The circularization step allows to obtain the full-length sequence of BCR and TCR, however, the primer used in the immunoreceptor enrichment step is located inside the constant region, hence the constant region is only partially sequenced. Yet, this partial sequence of the constant region is sufficient to identify BCR isotypes and detect isotype switching.
sCircle Single Cell Full-Length TCR/BCR Kit is available for human and mouse samples. The kit relies on poly-A capture followed by enrichment of the TCR/BCR regions using specific primers. For now, we have primers/kits with human- and mouse-specificity.
sCircle Single Cell Full-Length TCR/BCR Kit is available for TCR or BCR library preparation. Therefore, you will need to prepare two libraries: one for the transcriptome and one for the receptor (TCR or BCR).
One needs a concentration of 1.5 × 105 to 3.5 × 105 cells/mL (SD SCOPEchip). The desired viability of the cell suspension is >85%.
It is captured through the polyA tail of the mRNAs.
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