Count data, signal-to-noise ratios, and statistical analysis for TRBV12-3-TRBJ1-2-specific sequence reads obtained from spiked RNA samples. Results of linear regression analysis are indicated in the upper right region of the graph. Circles and triangles correspond to experimental replicates for each sample concentration. Normalized count data were then Log10 transformed. Count data for TRBV12-3-TRBJ1-2-specific sequence reads were normalized by subtracting the number of corresponding reads obtained for negative control samples consisting of unspiked PBMC RNA. Numbers along the X-axis indicate serial-diluted concentrations of spiked-in Jurkat RNA (by mass): 1 = 10% 2 = 1% 3 = 0.1% 4 = 0.01% 5 = 0.0.01. Correlation between concentration of spiked-in Jurkat RNA and number of TRBV12-3-TRBJ1-2-specific sequence reads. The experimental protocol was performed in replicate on PBMC RNA samples spiked at varying concentrations (10%, 1%, 0.1%, 0.01%, and 0.001%) with RNA obtained from a homogenous population of Leukemic Jurkat T cells (TRAV8-4-TRAJ3, TRBV12-3-TRBJ1-2 clonotype). The protocol generates indexed libraries that are ready for sequencing on Illumina platforms.Īssessing the sensitivity and reproducibility of the SMARTer approach Assessing the sensitivity and reproducibility of the SMARTer approach. Included in the kit are primers that incorporate Illumina-specific adaptor sequences during cDNA amplification. This kit leverages SMART technology and employs a 5' RACE-like approach to capture complete V(D)J variable regions of TCR transcripts. As the name suggests, the kit can be used to generate data for both alpha- and beta-chain diversity, either in the same experiment or separately. This kit is designed to work with a range of RNA input amounts depending on the sample type, and has been shown to generate high-quality libraries from as little as 10 ng to 3 μg of total RNA obtained from peripheral blood leukocytes, or from 50 to 10,000 purified T cells. licensing representative by e-mail at SMARTer Human TCR a/b Profiling Kit enables users to analyze T-cell receptor (TCR) diversity from input RNA samples.
#Tcr v beta repertoire license#
For license information, please contact a Takara Bio USA, Inc. patent application and/or pending claims of foreign counterparts. This product is sold under exclusive license from Ludwig Institute of Cancer Research, Ltd. This kit leverages our proven SMART technology ( Switching Mechanism at 5’ End of RNA Template) along with LNA technology to amplify TCR mRNA sequences in an unbiased manner with remarkable sensitivity. Libraries produced with the kit are indexed and ready for sequencing on Illumina platforms. In contrast with TCR profiling methods that involve PCR multiplexing, each round of library amplification utilizes a single primer pair for each TCR subunit. As the name suggests, the kit can be used to generate data for both TCR-alpha and TCR-beta chain diversity, either in the same experiment or separately. The kit employs a 5’ RACE-like approach to capture complete V(D)J variable regions of TCR transcripts, starting from human blood RNA samples (10 ng–3 µg) or directly from intact lymphocytes (50–10,000 purified T cells). The SMARTer Human TCR a/b Profiling Kit provides a powerful new solution for those seeking to perform T-cell receptor (TCR) repertoire analysis using NGS. # 635015 (48 rxns) are no longer available to order.
#Tcr v beta repertoire software#
Based on the same great chemistry, TCR v2 improves upon it with UMIs for confident clonotype calls, UDIs for increased multiplexing capabilities, and easy-to-use bioinformatics software for an end-to-end solution.Ĭat. For TCR sequencing, we recommend the upgraded version, the SMARTer Human TCR a/b Profiling Kit v2 (TCR v2).
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