Next-Generation Sequencing

Next-generation sequencing is a broad term for high-throughput, nucleic acid sequencing technologies. Our genomics core uses Illumina sequencing and Pacific Biosciences’ single molecule real-time sequencing.

The Illumina sequencing platform is based on reversible dye terminators, and can generate a massive amount of sequence information per run. It is the method of choice for most major sequencing centers. At full capacity, the HiSeq 2500 can produce more than 4 billion individual sequences from DNA samples in less than two weeks. It can also sequence whole human genomes or exomes, count transcripts in RNA samples, and identify genome-wide epigenetic signatures. Our genomics core currently operates four HiSeq 2500 instruments.

Illumina has released a new, personal sequencer–– the MiSeq––for low-throughput needs. The MiSeq will have a capacity of up to 15M reads per run, which it can complete in a day. We plan to use this technology for pilot projects and clinical genetic testing.

The Pacific Biosciences platform is exquisitely sensitive. It is able to continuously sequence a single molecule for up to 6 kilobases. The technology for this platform is based on recombinant polymerase and a cutting edge imaging system, which allows for visualization of individual base incorporation events during DNA synthesis. The platform is well suited for de-novo sequencing of small genomes and polishing of genomic assemblies for large genomes. Because the platform performs at the speed of the polymerase (2 bases per second), DNA samples are sequenced within hours instead of days. Rapid sequencing is of use when time is of the essence, as in a clinical testing or pathogen outbreak situation. Additionally, this platform allows unprecedented insight into the epigenetic organization of genomes beyond CpG methylation. The kinetic information obtained during sequencing allows modified bases such 5-methylcytosine, 5-hydroxymethylcytosine, 6-methyladenine, and 8-oxoguanine to be identified.

If you would like to sequence samples using our genomics core, you may submit your prepared libraries or we can prepare a DNA library for you. Guidance for experimental design is also available. Please see our sample submission page for our location and pricing information.