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.