NISC - NIH Intramural Sequencing Center Platforms

Platforms

NextGen Sequencing Technologies

NISC currently employs the following NextGen sequencing technologies:

Illumina NovaSeq 6000
NextSeq 550
Illumina MiSeq
PacBio Sequel II

Illumina NovaSeq 6000

The NovaSeq 6000 offers scalable sequencing through multiple flow cell modules. The most cost-effective module is the S4 which produces ~2.5 Tb per flow cell or ~ 625 Gb per lane. Due to the extremely high quantity of sequence output, this technology is primarily useful for projects requiring a very large number of reads, such as whole genome, whole exome and multiplexed transcriptome experiments.

Illumina NextSeq 550

The NextSeq 550 is a mid-range output sequencer. Multiple run options are offered, with outputs ranging from 16 Gb to 120 Gb and read lengths from 1 x 75 b to 2 x 150 b. This platform is ideal for smaller projects or those requiring non-standard run conditions, such as single-cell RNA-Seq.

Illumina MiSeq

MiSeq is a small-scale benchtop sequencer. This instrument is compatible with any of the Illumina libraries described above. MiSeq produces reads up to 300 bases in length from a paired-end read run (2x300 b) with a yield of ~10 Gb. This capability is primarily useful for sequencing microbial genomes and amplicons.

PacBio Sequel II

The PacBio Sequel II platform generates extremely long read lengths, while maintaining high consensus accuracy and unbiased coverage. The maximum read length is more than 50 kb and the average read length is 20 kb for whole genome samples. This platform can generate sequence reads that support high-quality assemblies of small (microbial) to intermediate size genomes and even human genomes. Other applications include amplicons, full length 16s sequencing, epigenomes, and RNA isoforms.For most applications one SMRT cell generates >50 Gb of data and completes in 20-30 hours. For amplicons and other library types with insert sizes < 10 kb, one SMRT cell generates > 3 million high-accuracy Circular Consensus Sequence (CCS) reads and completes in 10-30 hours.