Solid-phase amplification creates up to 1,000 identical copies of each single template molecule in close proximity with total densities on the order of more than one million single-molecule clusters per square millimeter.
Sequencing by Synthesis
Sequencing by synthesis (SBS) technology uses four fluorescently labeled nucleotides to simultaneously sequence the tens of millions of clusters on the flow cell surface. During each sequencing cycle, a single labeled deoxynucleoside triphosphate (dNTP) is added to the nucleic acid chain. The nucleotide label serves as a terminator for polymerization. After each dNTP incorporation, the fluorescent dye is imaged to identify the base and later enzymatically cleaved to allow incorporation of the next nucleotide. Since all four reversible terminator-bound dNTPs (A, C, T, G) are present as single separate molecules, natural competition minimizes incorporation bias. Base calls are made directly from signal intensity measurements during each cycle, thereby reducing raw error rates. The end result is highly accurate base-by-base sequencing that eliminates sequence-context specific errors, enabling robust base calling across the genome.
The TGC is home to an Illumina HiSeq 2500, an Illumina NextSeq 550 and an Illumina MiSeq. The HiSeq 2500 can be run in one of two modes: high output mode or rapid mode. In high output mode we offer sequencing runs of 50bp Single Read or Paired End, as well as 100bp Paired End. Researchers who submit two lanes for rapid run mode can have their samples sequenced at any length. The NextSeq can also be run in one of two mods: high output or mid output. NextSeq high output sequencing can yield 75, 150 or 300 cycles (one cycle = one base pair). In mid output sequencing 150 or 300 cycles are available. Researchers interested in MiSeq sequencing can receive 50, 150, 300, 500 or 600 cycles. For more information, please contact us.