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The immune repertoire (IR) refers to the sum of
all functionally diverse B cells and T cells in an individual's circulatory system at any given time.
B cells are an important component of the adaptive immune response, and their surface expresses B cell receptors (BCRs) that can recognize specific molecular patterns
in pathogens.
Understanding the BCRs map provides insight into adaptive immune responses not only in healthy individuals, but also in multiple diseases
.
Accurate identification of clones and isotypes expressed by the immune system provides a comprehensive understanding of the B cell population
.
Takara Bio's new SMART-Seq Human BCRs (with UMIs) provide a sensitive, accurate, and optimized BCR library analysis method
.
The kit combines SMART (Switching Mechanism at 5' end of RNA Template) full-length cDNA synthesis with 5'RACE technology to capture the intact V(D)J variable region of heavy (IgG/M/D/A/E) and light chain (IgK/L) genes for subsequent NGS analysis
.
The kit also includes a special molecular barcode (UMI) that eliminates PCR errors or duplication and sequencing bias, ensuring more accurate and reliable results
.
In addition, the use of double-ended special index (UDI, Unique Dual Index) can provide a complete experimental experience
from sample to library construction.
After sequencing, Cogent NGS Immune Profiler Software (Cogent IP) can be used to directly perform high-quality BCR profiling analysis of the Illumina sequencer's dismounted data, including clone identification and counting, and sequence analysis
of V(D)J regions.
Let's take a closer look at SMART-Seq Human BCRs (with UMIs).
Product features
◇ Complete BCR analysis—supports analysis of heavy chains (IgG, IgM, IgA, IgD, IgE) and light chains (IgK, IgL)
◇ Flexible sample start conditions—supports RNA initiation
from PBMCs (10 ng - 1 μg), B cells (1-100 ng), spleen or bone marrow (10 ng), whole blood (100 ng).
◇ Add UMI library to obtain more reliable sequencing data—UMI is introduced on SMARTer Oligo to correct sequencing errors and PCR bias
◇ Flexible sequencing platform selection—you can freely choose V(D)J full-length analysis, or only analyze CDR3
◇ Complete experimental and analytical workflow—with specially developed Cogent NGS Immune Profiler Software
Process summary
Figure 1 SMART-Seq Human BCR (with UMIs) technology flow
Product performance display
Fig.
2 Different RNA initiators can obtain clonotype detection data with high sensitivity and reproducibility
BCR was used to analyze IgA, IgD, IgG, IgM, IgK, and IgL libraries from 10 ng, 100 ng, and 1,000 ng human PBMC cell total RNA
.
Analyze sequencing data
using Cogent NGS Immune Profiler Software.
Therefore, if you want to carry out BCR receptor library analysis and pay attention to the sensitivity, specificity and reliability of clonal detection, but suffer from repeated optimization of experiments and no breakthrough in the article, this BCR analysis kit with molecular tags is worth trying!
|
Packaging volume |
Code No. |
SMART-Seq Human BCR (with UMIs) |
24 times |
634777 |
96 times |
634778 | |
4×96 times |
634776 |