Protocol for high-resolution multiplexed sequencing of single-cell full-length transcriptome with CBTi-seq.

Liyong He, Wenyi Zhang, Peidong Qi, Zhe Jiao, Dongmei Zhang, Xiangwei Zhao

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Article in STAR protocols, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

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5 · Who and what money

Authors and funding

6 authors.

Liyong HeMedical Research Center, Jiangsu Provincial Medical Key Discipline (Laboratory) Cultivation Unit of Immunology, Nantong First People's Hospital, Southeast University, Nantong, Jiangsu, China; State Key Laboratory of Digital Medical Engineering, School of Biological Science & Medical Engineering, Southeast University, Nanjing 211196, China. Electronic address: heliyong@seu.edu.cn.

Wenyi ZhangState Key Laboratory of Digital Medical Engineering, School of Biological Science & Medical Engineering, Southeast University, Nanjing 211196, China.

Peidong QiState Key Laboratory of Digital Medical Engineering, School of Biological Science & Medical Engineering, Southeast University, Nanjing 211196, China.

Zhe JiaoState Key Laboratory of Digital Medical Engineering, School of Biological Science & Medical Engineering, Southeast University, Nanjing 211196, China.

Dongmei ZhangMedical Research Center, Jiangsu Provincial Medical Key Discipline (Laboratory) Cultivation Unit of Immunology, Nantong First People's Hospital, Southeast University, Nantong, Jiangsu, China. Electronic address: zdm_ntyy@163.com.

Xiangwei ZhaoState Key Laboratory of Digital Medical Engineering, School of Biological Science & Medical Engineering, Southeast University, Nanjing 211196, China; Department of Thoracic Surgery, Nantong First People's Hospital, Southeast University, Nantong, Jiangsu, China. Electronic address: xwzhao@seu.edu.cn.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Balancing throughput and full-length coverage remains challenging in single-cell RNA sequencing (scRNA-seq). Here, we present combinational barcoded Tn5 transposon insertion sequencing (CBTi-seq), a highly scalable protocol for constructing multiplexed, full-length transcriptome libraries from single cells and micro-region tissues. The workflow details sample acquisition, one-step reverse-transcription PCR (RT-PCR) cDNA amplification, orthogonal combinational barcoded Tn5 tagmentation, multiplexed pooling, dual-option purification, and final library enrichment. For complete details on the use and execution of this protocol, please refer to He et al.

Indexed as

Cell-based AssaysCell BiologyGenomicsMolecular BiologyMolecular/Chemical ProbesRNAseqSequencingSingle Cell

Identifiers

PMID42545848

DOI10.1016/j.xpro.2026.104765

PMCPMC13469961

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.