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.

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