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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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RNA Stability01:53

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Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
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RNA Structure01:23

RNA Structure

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Overview
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
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Alternative RNA Splicing02:18

Alternative RNA Splicing

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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
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Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

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Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
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Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

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Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
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Nuclei Isolation from Fresh Frozen Brain Tumors for Single-Nucleus RNA-seq and ATAC-seq
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Single-Cell RNA-Seq by Multiple Annealing and Tailing-Based Quantitative Single-Cell RNA-Seq (MATQ-Seq).

Kuanwei Sheng1, Chenghang Zong2

  • 1Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.

Methods in Molecular Biology (Clifton, N.J.)
|April 28, 2019
PubMed
Summary

Multiple annealing and dC-tailing-based quantitative single-cell RNA-sequencing (MATQ-seq) offers high accuracy and sensitivity for detecting transcriptome heterogeneity. This method achieves ~90% capture efficiency and analyzes total RNA, including nonpolyadenylated transcripts.

Keywords:
BiotechnologyLibrary preparationRNA-seqSingle cellTotal RNATranscriptomics

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Single-cell technologies are crucial for analyzing biological processes at high resolution.
  • Current single-cell RNA sequencing (scRNA-seq) methods require improved accuracy and sensitivity to detect subtle transcriptome variations.
  • Detecting nonpolyadenylated transcripts remains a challenge in existing scRNA-seq assays.

Purpose of the Study:

  • To introduce a novel single-cell RNA sequencing method, MATQ-seq, with enhanced performance.
  • To improve the capture efficiency and sensitivity of single-cell transcriptomic analysis.
  • To enable the detection of both polyadenylated and nonpolyadenylated RNA molecules in single cells.

Main Methods:

  • Development of the multiple annealing and dC-tailing-based quantitative single-cell RNA sequencing (MATQ-seq) protocol.
  • Application of MATQ-seq for comprehensive transcriptome profiling at the single-cell level.
  • Quantification of RNA capture efficiency and sensitivity using the MATQ-seq assay.

Main Results:

  • MATQ-seq demonstrated a high RNA capture efficiency of approximately 90%.
  • The method provides high accuracy and sensitivity for detecting transcriptome heterogeneity.
  • MATQ-seq successfully detected nonpolyadenylated transcripts, expanding the scope of single-cell analysis.

Conclusions:

  • MATQ-seq represents a significant advancement in single-cell RNA sequencing technology.
  • The method enhances the ability to study cellular heterogeneity through improved transcriptomic analysis.
  • MATQ-seq offers a more comprehensive approach to single-cell RNA profiling by including nonpolyadenylated transcripts.