Instruct: Identify the main category of Biorxiv papers based on the titles and abstracts
Query: An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences T cell receptor (TCR) repertoires can be profiled using next generation sequencing (NGS) to monitor dynamical changes in response to disease and other perturbations. Several strategies for profiling TCRs have been recently developed with different benefits and drawbacks. Genomic DNA-based bulk sequencing, however, remains the most cost-effective method to profile TCRs. The major disadvantage of this method is the need for multiplex target amplification with a large set of primer pairs with potentially very different amplification efficiencies. One approach addressing this problem is by iteratively adjusting the concentrations of the primers based on their efficiencies, and then computationally correcting any remaining bias. Yet there are no standard, publicly available protocols to process and analyze raw sequencing data generated by this method. Here, we utilize an equimolar primer mixture and propose a single statistical normalization step that efficiently corrects for amplification bias post sequencing. Using samples analyzed by both approaches, we show that the concordance between bulk clonality metrics obtained from using the commercial kits and that developed herein is high. Therefore, we suggest the method presented here as an inexpensive and non-commercial alternative for measuring and monitoring adaptive dynamics in TCR clonotype repertoire.
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The categories of this paper are: plant
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The categories of this paper are: animal
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The categories of this paper are: scientific
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The categories of this paper are: pharmacology
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The categories of this paper are: immunology
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The categories of this paper are: genomics
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The categories of this paper are: biochemistry
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The categories of this paper are: education
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The categories of this paper are: genetics
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The categories of this paper are: cell
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The categories of this paper are: paleontology
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The categories of this paper are: physiology
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The categories of this paper are: synthetic
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The categories of this paper are: and
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The categories of this paper are: molecular
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The categories of this paper are: bioengineering