Chemistry and imaging on MiSeq

Four-Channel Sequencing by Synthesis (SBS) Chemistry
In four-channel SBS, bases are identified using four different fluorophores for each base and four images per sequencing cycle.
  • The sequencing cycle begins with a chemistry step where all four deferentially labeled bases are added to the flow cell.
  • Following nucleotide incorporation, the imaging cycle begins and includes the capture of four distinct images using four different wavelength bands.
  • The images are processed with image analysis software to determine which nucleotides were incorporated at each cluster position across the flow cell.
  • With four-channel sequencing, every sequencing cycle requires four dyes and four images to determine the DNA sequence.
The MiSeq and HiSeq Series Systems currently use four-channel SBS.
For more information, see SBS Technology Spotlight.
For any feedback or questions regarding this article (Illumina Knowledge Article #3757), contact Illumina Technical Support [email protected].
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