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Products >  cDNA_Synthesis_and_Library_Construction >  NGS_Automation >  Single-Cell_Automation >  High_Throughput_Human_TCR_Profiling

Human TCR Profiling and Differential Expression on the SMARTer ICELL8 Single-Cell System

Our high-throughput TCR profiling solution leverages SMART technology (Switching Mechanism At 5’ End of RNA Template) and employs a 5’ RACE-like approach to capture complete V(D)J variable regions of TCR transcripts from hundreds of single cells isolated on the SMARTer ICELL8 systems. As the name suggests, the workflow allows generation of libraries that provide information on both alpha- and beta-chain diversity. The workflow incorporates Illumina-specific adaptor sequences during cDNA amplification for the generation of indexed libraries that are ready for sequencing on Illumina platforms. The application also enables you to perform differential expression analysis by 5’ end capture in parallel.

Single T cells isolated with the SMARTer ICELL8 system, as well as positive and negative control samples, are dispensed into a 5,184-nanowell TCR chip that contains preprinted barcodes. Each SMARTer ICELL8 TCR Chip (coming soon) contains 1,728 unique barcodes, and each barcode is printed three times on the chip. Following identification of single-cell-bearing wells using CellSelect Software, SMART RT-PCR reagents are dispensed to the wells of samples chosen for further processing. Barcoded cDNA from the selected wells are PCR-amplified in-chip. The full-length barcoded amplicons are then pooled off-chip, and the purified cDNA is used as template in the first TCR-specific PCR. The product from the first TCR-specific PCR is used as a template in a second TCR-specific PCR to incorporate Illumina indexed primers.

  At-A-Glance   Documents

Features

  • Sensitivity—RACE-based approach allows for the detection of low-abundance TCR variants
  • Specificity—Full-length reads, with majority of reads on target, and accurate pairing information
  • Differential expression by 5’ sequence enrichment—generate differential expression data for all transcripts by mapping the 5’ ends
  • Flexible, high-throughput single-cell analysis—generate sequencing data from hundreds of single cells in parallel and analyze cells from up to 8 different samples in a single experiment

Applications

  • Human TCR repertoire analysis in single cells (TCRa and TCRb clonotypes)
  • Differential expression by 5’ end capture

Components

The following items are required to complete the workflow:

  • SMARTer ICELL8 TCR Chip (Coming soon; Cat. No. 640178)
  • SMARTer ICELL8 Human TCR a/b Profiling Reagent Kit (Cat. No. 640182)
  • SMARTer ICELL8 Collection Kit (Cat. No. 640048)
  • SMARTer ICELL8 Loading Kit (Cat. No. 640109)
  • SMARTer ICELL8 Human TCR a/b Profiling - Indexing Primer Set (Cat. Nos. 640179, 640180 & 640181)*

*Note: Cat. No. 640179 contains enough primers for one chip. If you plan on processing multiple chips at a time, please purchase the appropriate indexing primer set. Cat. No. 640180 has indexing primers for up to five chips, while Cat. No. 640181 has primers for up to 10 chips.

Additional Product Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


 
 
Products
New Cat. # Product Contents Size Price License Units Select
  640109 SMARTer™ ICELL8® Loading Kit 1 Pkg $25.00  
  640048 SMARTer™ ICELL8® Collection Kit 1 Pkg $25.00  
640179 SMARTer® ICELL8® Human TCR a/b Profiling - Indexing Primer Set 1 Chip $198.00  
640180 SMARTer® ICELL8® Human TCR a/b Profiling - Indexing Primer Set 5 Chips $648.00  
640181 SMARTer® ICELL8® Human TCR a/b Profiling - Indexing Primer Set 10 Chip $1,250.00  
640182 SMARTer® ICELL8® Human TCR a/b Profiling Reagent Kit Each $499.00 License Statements
 

 

Required Products
Cat. # Product Contents Size Price # of Units Select
640000 SMARTer™ ICELL8® Single-Cell System 1 each Call
 

 

 
 

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