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Related Concept Videos

Next-generation Sequencing03:00

Next-generation Sequencing

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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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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. 
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DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
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Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
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Related Experiment Video

Updated: Aug 3, 2025

Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing
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Nuclei on the Rise: When Nuclei-Based Methods Meet Next-Generation Sequencing.

Tamer Butto1, Kanak Mungikar2, Peter Baumann3,4

  • 1Institute for Pharmaceutical and Biomedical Sciences, Johannes Gutenberg-University, 55128 Mainz, Germany.

Cells
|April 13, 2023
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Summary

Nuclei-based studies, especially using next-generation sequencing, offer insights into cell populations but present challenges. Understanding these methods is key for accurate transcriptomic analysis.

Keywords:
cell-type-specific isolationepigeneticsnext-generation sequencingnuclei isolationsingle-cell sequencingsingle-nucleus sequencingtranscriptomics

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Area of Science:

  • Molecular Biology
  • Genomics
  • Cell Biology

Background:

  • Nuclei-based studies have surged in the last decade, driven by advances in next-generation sequencing.
  • These methods enable the analysis of molecular states within heterogeneous cell populations.
  • Increasingly affordable sequencing and optimized nuclei isolation techniques facilitate these studies.

Purpose of the Study:

  • To provide a comprehensive overview of the rise of nuclei-based studies.
  • To critically evaluate the advantages and disadvantages of nuclei-based approaches.
  • To focus on the utility of these methods for transcriptomic sequencing analyses.

Main Methods:

  • Review of recent literature on nuclei-based studies.
  • Analysis of methodologies for nuclei isolation and selection.
  • Evaluation of transcriptomic sequencing applications using nuclei.

Main Results:

  • Nuclei-based studies offer powerful new methods for molecular profiling.
  • Key challenges associated with these techniques require careful consideration.
  • Specific experimental designs impact the quality of transcriptomic data.

Conclusions:

  • Nuclei-based studies are valuable for understanding cellular heterogeneity.
  • Critical assessment of methods and experimental design is essential for reliable transcriptomic insights.
  • Appropriate application of these strategies yields biologically accurate information.