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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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Protocol for optimized nasal mucosa sample processing to obtain high-quality scRNA-seq and scATAC-seq data.

Yaling Huang1, Yisha Wu2, Shikai Han2

  • 1BGI Research, Shenzhen 518083, China.

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|September 8, 2024
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Summary

This study presents a new protocol for analyzing human nasal mucosa. It enables high-quality single-cell RNA sequencing and single-cell ATAC sequencing from the same tissue sample for disease research.

Keywords:
Cell isolationCell separation/fractionationGenomicsSequencingSingle Cell

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Area of Science:

  • Biomedical Research
  • Genomics
  • Molecular Biology

Background:

  • Nasal mucosa examination is vital for diagnosing nasal cavity diseases.
  • Simultaneous high-quality single-cell RNA sequencing (scRNA-seq) and single-cell ATAC sequencing (scATAC-seq) from nasal mucosa is technically challenging.
  • Existing methods may not efficiently capture both transcriptomic and epigenomic data from these sensitive tissues.

Purpose of the Study:

  • To develop and present a robust protocol for processing human nasal mucosa.
  • To enable the simultaneous generation of high-quality scRNA-seq and scATAC-seq data from the same samples.
  • To facilitate comprehensive research into nasal cavity diseases at the single-cell level.

Main Methods:

  • The protocol involves meticulous steps for human nasal mucosa tissue extraction.
  • It includes optimized mechanical and enzymatic dissociation techniques.
  • Procedures cover red blood cell lysis, viability assessment, and library preparation for both scRNA-seq and scATAC-seq, followed by quality control.

Main Results:

  • The presented protocol successfully yields high-quality data for both scRNA-seq and scATAC-seq from human nasal mucosa.
  • The method ensures the integrity of both transcriptomic and epigenomic information.
  • Quality control metrics confirm the suitability of the prepared libraries for downstream analysis.

Conclusions:

  • This protocol provides a standardized and effective method for multi-omic single-cell analysis of nasal mucosa.
  • It overcomes previous limitations in obtaining both transcriptomic and epigenomic data simultaneously.
  • This advancement will significantly aid in the detailed investigation and diagnosis of nasal cavity diseases.