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Related Concept Videos

RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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Related Experiment Video

Updated: Jan 11, 2026

Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues
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High-Resolution Multiplexed Sequencing of Single-Cell Full-length Transcriptome Via Combinational Barcoded Tn5

Liyong He1, Kaitong Dang1, Qian Sun2

  • 1State Key Laboratory of Digital Medical Engineering, School of Biological Science & Medical Engineering, Southeast University, Nanjing, 211189, China.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|November 11, 2025
PubMed
Summary
This summary is machine-generated.

Combinational Barcoded Tn5 Transposon Insertion sequencing (CBTi-seq) offers high-resolution, full-length transcriptome sequencing in single cells. This cost-effective method accurately identifies splice variants and transcript abundance, advancing single-cell analysis.

Keywords:
alternative splicingbarcoded Tn5 transposasefull‐length transcriptomemultiplexedsingle cellspermatogenesis

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Single-cell transcriptome analysis is advancing rapidly in depth and breadth.
  • A key challenge is balancing cell throughput with full-length transcript coverage.
  • Existing methods often struggle with resolution, cost, or time.

Purpose of the Study:

  • To introduce CBTi-seq, a novel method for high-resolution, full-length single-cell transcriptome sequencing.
  • To demonstrate CBTi-seq's ability to accurately reconstruct splice variants and structural variations.
  • To provide a cost-effective and time-efficient alternative to current sequencing methods.

Main Methods:

  • CBTi-seq utilizes Tn5 transposase for molecular assembly of combinatorial barcodes and unique molecular identifiers (UMIs).
  • End-to-end sequencing enables base-pair precision reconstruction of transcripts.
  • Orthogonal barcode design enhances multiplexing flexibility, while Tn5-delivered UMIs reduce bias and improve quantification.

Main Results:

  • CBTi-seq achieves superior sensitivity and resolution compared to commercial methods.
  • The method significantly reduces costs and processing time (approximately 5 hours).
  • Robust identification of cell-type-specific alternative splicing and isoform switching was demonstrated in gene-edited and human testicular cells.

Conclusions:

  • CBTi-seq provides a powerful tool for high-resolution single-cell transcriptome analysis.
  • The method facilitates the study of complex biological processes like spermatogenesis.
  • CBTi-seq has potential applications in reproductive development research and diagnostics.