Single-cell Rapid Capture Hybridization sequencing reliably detects isoform usage and coding mutations in targeted genes.

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Tác giả: Noorul Amin, Natasha S Anstee, Chong Chyn Chua, Nadia M Davidson, David C S Huang, Jafar S Jabbari, Hongke Peng, Matthew E Ritchie, Andrew W Roberts, Rachel Thijssen, Luyi Tian, Changqing Wang, Andrew H Wei, Yupei You

Ngôn ngữ: eng

Ký hiệu phân loại: 598.635 *Bonasa

Thông tin xuất bản: United States : Genome research , 2025

Mô tả vật lý:

Bộ sưu tập: NCBI

ID: 700706

Single-cell long-read sequencing has transformed our understanding of isoform usage and the mutation heterogeneity between cells. Despite unbiased in-depth analysis, the low sequencing throughput often results in insufficient read coverage, thereby limiting our ability to perform mutation calling for specific genes. Here, we developed a single-cell Rapid Capture Hybridization sequencing (scRaCH-seq) method that demonstrates high specificity and efficiency in capturing targeted transcripts using long-read sequencing, allowing an in-depth analysis of mutation status and transcript usage for genes of interest. The method includes creating a probe panel for transcript capture, using barcoded primers for pooling and efficient sequencing via Oxford Nanopore Technologies platforms. scRaCH-seq is applicable to stored and indexed single-cell cDNA, which allows analysis to be combined with existing short-read RNA-seq data sets. In our investigation of
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