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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.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
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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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Sanger Sequencing01:57

Sanger Sequencing

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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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Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
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Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

10.5K
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
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Updated: May 3, 2026

Pyrosequencing: A Simple Method for Accurate Genotyping
13:06

Pyrosequencing: A Simple Method for Accurate Genotyping

Published on: January 8, 2008

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Pyrosequencing: powerful and quantitative sequencing technology.

Martin Kreutz1, Norbert Hochstein1, Julia Kaiser1

  • 1Qiagen GmbH, Hilden, Germany.

Current Protocols in Molecular Biology
|February 11, 2014
PubMed
Summary
This summary is machine-generated.

Pyrosequencing offers a versatile DNA analysis method for various applications. This workflow details sample-to-result procedures for methylation analysis, SNP genotyping, and allele quantification.

Keywords:
AQDNA methylationSNPgenome analysispyrosequencingsequencing

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Pyrosequencing for Microbial Identification and Characterization
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Last Updated: May 3, 2026

Pyrosequencing: A Simple Method for Accurate Genotyping
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Pyrosequencing for Microbial Identification and Characterization
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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Pyrosequencing is a key sequencing-by-synthesis technology.
  • It enables diverse DNA analysis applications beyond de novo sequencing.
  • Quantitative analysis of genomic features is crucial in molecular diagnostics.

Purpose of the Study:

  • To present a comprehensive pyrosequencing workflow.
  • To detail applications in methylation analysis, SNP genotyping, and allele quantification.
  • To provide a protocol for bisulfite treatment essential for methylation analysis.

Main Methods:

  • Description of a complete sample-to-result pyrosequencing workflow.
  • Inclusion of a bisulfite treatment protocol for cytosine conversion.
  • Adaptation of the workflow for quantitative DNA analyses.

Main Results:

  • A standardized workflow suitable for multiple pyrosequencing applications.
  • Successful implementation of quantitative analyses including methylation, SNPs, and allele ratios.
  • A validated protocol for bisulfite treatment enhancing methylation analysis accuracy.

Conclusions:

  • Pyrosequencing provides a robust platform for quantitative DNA analysis.
  • The described workflow streamlines applications in epigenetics and genetic variation studies.
  • Optimized bisulfite treatment is critical for reliable methylation quantification using pyrosequencing.