Advances in Genome Biology and Technology (AGBT) General Meeting
Solutions for NGS analysis of ultra-low inputs and degraded samples
The AGBT General Meeting brings together experts in nucleic acid sequencing and genomic studies for discussions on the latest advancements in these technologies and their range of applications. We enjoyed spending time with and learning from our colleagues (and meeting new ones!) at AGBT 2019 in Florida.
We're looking forward to seeing you at AGBT 2020. In the meantime, we invite you to review our materials from past AGBT meetings and reach out to us with any questions or requests via the "speak with us" link below.
AGBT 2019: Women's Networking Event
Supporting and promoting the advancement of women in science is important to our company. Takara Bio is proud to have hosted the 2019 Women's Networking Event at AGBT, and we thank our attendees for a wonderful evening! Read more about our spotlighted employees below.
AGBT 2018: talk and poster
High-throughput single-cell T-cell receptor profiling by SMART technology
Single-cell T-cell receptor (scTCR) clonotype analysis permits the determination of the specific TCR alpha-beta (α/β) chain pairing expressed on each cell. This pairing information allows researchers to gain insight into T-cell heterogeneity and plasticity, determine the contribution of the pairing to antigen specificity of the individual TCR, and design therapeutic antibodies. Here we employ a 5'-RACE-like approach and SMART technology, in conjunction with two novel next-generation sequencing (NGS) library preparation kits, using the same primer pairs, to capture full-length variable regions of TCR-α and -β chains.
Method 1, using the SMARTer Human scTCR a/b Profiling Kit, permits NGS library preparation of FACS-sorted cells in 96-well plates. We present data showing α/β pairing from Jurkat, CCRF, PBMCs, and CD4+ T cells. In addition to the sensitivity of this method, the ability to pool the cDNA from 96 wells into 12 sequencing libraries adds to the ease of use. Consistent with immunology reports, unstimulated CCRF-CEM cells examined with this kit expressed a TCR-β but not a TCR-α chain.
Method 2 is an adaptation of the process above that scales to ~1,200 single cells using the SMARTer ICELL8 Single-Cell System, which enables single-cell isolation and nanoliter PCR in a nanowell chip. For proof-of-principle studies, Jurkat cells and CCRF-CEM cells were processed using an ICELL8 chip preprinted with barcoded oligos. Paired TCR α/β Jurkat clonotypes were detected in 77% and 87% cells in mixed and single cell populations, respectively.
The ability of the core biochemistry and PCR components of these kits to be used with either FACS-sorted cells in 96-well plates or >1,000 cells in novel ICELL8 chips points to the general utility and scalability of this approach in understanding paired scTCR clonotype diversity.
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