Detection of ultra-rare mutations by next-generation sequencing

Detection of ultra-rare mutations by next-generation sequencing

September 4, 2012 | vol. 109 | no. 36 | Michael W. Schmitt, Scott R. Kennedy, Jesse J. Salk, Edward J. Fox, Joseph B. Hiatt, and Lawrence A. Loeb
The article introduces a novel method called Duplex Sequencing, which aims to improve the accuracy of next-generation DNA sequencing by reducing the error rate associated with the current technology. The method involves tagging both strands of a DNA duplex with complementary double-stranded nucleotide sequences, allowing for the independent tagging and sequencing of each strand. This approach significantly reduces the background error rate, making it possible to detect rare variants in genetically heterogeneous mixtures, such as tumors or mixed microbial populations. The authors demonstrate that Duplex Sequencing has a theoretical background error rate of less than one artifactual mutation per billion nucleotides sequenced. They also show that the method can be used to identify sites of DNA damage by comparing the sequences from complementary strands of a DNA duplex. The technique is validated using M13mp2 DNA and human mitochondrial DNA, demonstrating its potential for high-sensitivity detection of rare mutations and accurate molecular counting.The article introduces a novel method called Duplex Sequencing, which aims to improve the accuracy of next-generation DNA sequencing by reducing the error rate associated with the current technology. The method involves tagging both strands of a DNA duplex with complementary double-stranded nucleotide sequences, allowing for the independent tagging and sequencing of each strand. This approach significantly reduces the background error rate, making it possible to detect rare variants in genetically heterogeneous mixtures, such as tumors or mixed microbial populations. The authors demonstrate that Duplex Sequencing has a theoretical background error rate of less than one artifactual mutation per billion nucleotides sequenced. They also show that the method can be used to identify sites of DNA damage by comparing the sequences from complementary strands of a DNA duplex. The technique is validated using M13mp2 DNA and human mitochondrial DNA, demonstrating its potential for high-sensitivity detection of rare mutations and accurate molecular counting.
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