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AAT Bioquest

What are the examples of short read sequencing technologies?

Posted June 4, 2024


Answer

These are some examples of short-read sequencing technologies:

Illumina: In this SRS technology, single-stranded DNA-binding proteins are first used to amplify DNA. The amplified DNA template is then bridged by the addition of fluorescent-labeled deoxynucleoside triphosphates. 

454 pyrosequencing: This SRS technology uses emulsion PCR to amplify DNA within individual water-in-oil droplets, preventing unwanted reactions between similar sequences. This process creates microbead-bound DNA clones, with each bead carrying multiple copies of a single DNA sequence. DNA polymerase is added to facilitate replication, and the incorporation of nucleotides is tracked by monitoring pyrophosphate release, which serves as a signal indicating successful nucleotide addition. 

Ion Torrent: Ion Torrent sequencing involves clonal amplification via emulsion PCR, which generates microbead-bound DNA clones. The DNA clones are trapped on microbeads. DNA polymerase is then added and the nucleotides are washed.  The incorporation of deoxynucleoside triphosphate is monitored by detecting the release of protons with a pH sensor. As deoxynucleoside triphosphates are added to the DNA strand, the release of protons changes the pH, allowing precise sequencing.

SOLiD: SOLiD sequencing uses emulsion PCR to amplify DNA templates bound to microbeads. These templates are flanked by adapters and hybridized to the growing complementary strand using DNA ligase.

Additional resources

Mapping short DNA sequencing reads and calling variants using mapping quality scores

Next Generation Sequencing (NGS)

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