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Related Concept Videos

Sanger Sequencing01:57

Sanger Sequencing

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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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.
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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. 
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Maxam-Gilbert Sequencing01:05

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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.
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Related Experiment Video

Updated: Jul 15, 2026

Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo
10:51

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DNA sequencing with a hexamer string primer and dye-labeled terminators

W Hou1, L M Smith

  • 1Department of Chemistry, University of Wisconsin-Madison 53706.

Analytical Biochemistry
|August 15, 1994
PubMed
Summary

This study introduces a novel hexamer string primer for DNA sequencing, enhancing signal intensity. This method achieves high accuracy and read lengths comparable to standard primers.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Fluorescence-based DNA sequencing is a cornerstone of genetic analysis.
  • Optimizing primer strategies is crucial for improving sequencing efficiency and accuracy.
  • Current methods face limitations in signal strength and read length.

Purpose of the Study:

  • To develop and evaluate a novel hexamer string primer for fluorescence-based DNA sequencing.
  • To assess the impact of this primer on signal intensity, read length, and base-calling accuracy.
  • To compare the performance of the hexamer string primer against a standard 18-mer primer.

Main Methods:

  • Development of a fluorescence-based DNA sequencing protocol utilizing a hexamer string primer.
  • Employing dye-labeled terminators for nucleotide incorporation.
  • Sequencing of single-stranded M13mp18 DNA and double-stranded M13mp19 DNA.
  • Analysis of read length, base-calling accuracy, and signal intensity.

Main Results:

  • Achieved an average read length of 393 bases with 99.6% accuracy for single-stranded M13mp18 DNA.
  • Obtained an average read length of 367 bases with 99.4% accuracy for double-stranded M13mp19 DNA.
  • Demonstrated stronger signal intensity compared to a standard 18-mer primer.
  • Maintained comparable sequence resolution and readable sequence length.

Conclusions:

  • The hexamer string primer offers a viable alternative for fluorescence-based DNA sequencing.
  • This method provides significant improvements in signal intensity without compromising read length or accuracy.
  • The findings support the utility of hexamer string primers for enhanced DNA sequencing applications.