DNA sequencing is the process of determining the order of nucleotides in fragments of DNA. Several sequencing techniques have been evolved over the past years.[1-3] The most commonly used today are based on sequencing-by-synthesis (SBS) such as Reversible termination Sequencing or Single molecule real time (SMRT) Sequencing. The fundamental difference between these techniques is the type of nucleotide analog incorporated by a specific polymerase to produce the observable output.
| Sequencing technique | Type of nucleotide |
|---|---|
| Reversible termination sequencing | 3‘-OH reversible blocked dNTPs (NRTs) |
| Single molecule real time (SMRT) Sequencing | zeta-phosphate-labeled dNTPs (d6NPs) |
| Chain termination sequencing | (labeled) ddNTPs |
Auxiliary reagents: Resolution enhancing nucleotides
[1] Chen et al. (2014) DNA Polymerases Drive DNA Sequencing-by-Synthesis Technologies: Both Past and Present. Front. Microbiol. 5:1.
[2] Heather and Chain (2016) The sequence of sequencers: The history of sequencing DNA. Genomics 107(1):1.
[2] Hu et al. (2021) Next-generation sequencing technologies: An overview. Hum Immunol 82(11):801.
