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Related Concept Videos

Next-generation Sequencing03:00

Next-generation Sequencing

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.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
RNA-seq03:21

RNA-seq

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 microarray-based...
Modern Molecular Taxonomy01:29

Modern Molecular Taxonomy

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: Jun 6, 2026

Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease
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Published on: April 4, 2018

Overcoming methodical limits of standard RHD genotyping by next-generation sequencing.

S Stabentheiner1, M Danzer, N Niklas

  • 1Red Cross Transfusion Service for Upper Austria, Krankenhausstrasse 7, Linz, Austria. stephanie.stabentheiner@o.roteskreuz.at

Vox Sanguinis
|December 8, 2010
PubMed
Summary

Next-generation sequencing, specifically pyrosequencing, offers a powerful new method for RHD genotyping, accurately detecting RHD gene variations missed by standard Sanger sequencing. This advancement improves the molecular characterization of complex RhD phenotypes.

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Last Updated: Jun 6, 2026

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Area of Science:

  • Genetics
  • Molecular Biology
  • Immunology

Background:

  • Molecular variations in the RHD gene can lead to reduced D antigen expression and altered Rh phenotypes, often challenging standard serological typing.
  • Sequence-based typing is the definitive method for identifying rare and unknown RHD genotypes.

Purpose of the Study:

  • To compare the efficacy of standard Sanger sequencing with a novel next-generation sequencing (NGS) pyrosequencing approach for RHD genotyping.
  • To evaluate the mutation detection capabilities of both sequencing methods in samples with weak RhD phenotypes.

Main Methods:

  • Twenty-six DNA samples with weak RhD reactions were analyzed using both Sanger sequencing and NGS pyrosequencing.
  • Sequence analysis covered the complete coding region and adjacent intronic sequences of the RHD gene.

Main Results:

  • Sanger sequencing identified 39 RHD polymorphisms in 21 samples.
  • NGS pyrosequencing detected all but two alterations, achieving a 94.9% concordance rate and resolving complex mutations and duplications.
  • NGS provided superior resolution for cis/trans linkage and characterization of a 37 bp duplication.

Conclusions:

  • NGS pyrosequencing represents a significant advancement for high-throughput, clonal molecular RHD genotyping.
  • Further methodological refinement is necessary before NGS can be implemented as a routine diagnostic service.