Illumina Sequencing Services

Depending on the project size and specific applications, ORB will perform sequencing using the NextSeq 500, HiSeq 2500, and HiSeq 4000 instruments. See Table I to view specifications for each of the three sequencers and the most appropriate applications for each.

Single-molecule sequencing technologies from Pacific Biosciences, Oxford Nanopore, and other companies, that enable reads of several thousand base-pairs have recently arrived on the market. However, instruments based on these newer technologies cannot match the combination of accuracy, quantity, and length of reads produced by the Illumina sequencing instruments. ORB expects that Illumina will continue to dominate RNA sequencing for the next few years.

ORB provides complete solution genetic, transcriptomic, metatranscriptomic, and epigenetic-based services using Illumina sequencing technology, including comprehensive data analysis with all sequencing services. Each sequencing project is customized to take full advantage of Illumina's multiplexing capability and minimize per sample cost. Follow the links below to learn more about ORB’s core and focused Illumina sequencing services.


RNA Sequencing Routinely ORB partners with researchers worldwide in order to execute challenging RNA sequencing applications such as sequencing of RNA from immunoprecipitated RNA-protein complexes, small RNA sequencing from ultra-low input material, and biomarker discovery initiatives querying for long non-coding, mRNA, and small RNA markers associated with the presence or progression of disease as well as the efficacy of therapeutic agents.

Long RNA Sequencing Examine gene expression patterns, alternative splicing isoforms, and long non-coding RNA using library preparation kits from industry leading manufacturers such as Illumina, NuGen, and Clontech.

Small RNA Sequencing ORB offers extensively customizable small RNA sequencing services to interrogate numerous small RNA biotypes, e.g. mature microRNA, pre-microRNA, piRNA, snRNA, snoRNA, rRNA, tRNA, etc.

Oncopanels Targeted oncogene analysis facilitates the evaluation of germline and somatic mutations, insertions and deletions (INDELs), single nucleotide polymorphisms (SNPs), structural variants, copy number, loss of heterozygosity, chromosomal and short tandem repeats.

Metatranscriptomics Profile gene expression signatures of microbial populations from in vivo samples, such as feces, to identify relationships between gut microflora and pathological conditions.

Cell-Free DNA Bisulfite Sequencing Analyze circulating methylation patterns from cell-free DNA extracted from blood-based biofluid samples to identify epigenetic biomarkers.

Exosomes ORB provides sequencing services to investigate genetic information, like long and small RNA molecules contained within exosomes that present in biofluid samples.

Biofluids ORB offers multiple sequencing services to interrogate RNA and DNA patterns from a wide variety of biological fluids such as whole blood, plasma, serum, saliva, bronchial lavage, urine, milk, semen, tears as well as cerebrospinal, amniotic, lymphatic, ascites, and synovial fluids.
 

Table I. Sequencing instrument, metrics, and typical applications at ORB.
Sequencing instrument Standard configurations Run Time (H)1 Reads total2
| per lane (M)
Typical applications
HiSeq 2500 1 x 50 bp 36 2000 | 250 Small RNA, methylation
2 x 50 bp 60 " mRNA, metatranscriptome
2 x 100 bp 120 " Alternative splicing, non-coding RNA
NextSeq 5003 1 x 75 bp 11 400 | NA Small RNA, mRNA
2 x 75 bp 18 " mRNA, metatranscriptome, alternative splicing, ncRNA
2 x 150 bp 30 " RNA polymorphisms, exome
HiSeq 4000 1 x 50 bp 15 2500 | 313 Methylation
2 x 75 bp 44 " mRNA, metatranscriptome, alternative splicing, ncRNA
2 x 150 bp 84 " RNA polymorphisms, exome, whole genome
NovoSeq 60004 2 x 50 bp 19 3000 | NA mRNA, metatranscriptome
2 x 100 bp 29 " Alternative splicing, non-coding RNA
2 x 150 bp 40 " RNA polymorphisms, exome, whole genome

1. Run times in hours are estimated for HiSeq 2500 and 4000 using Illumina specifications which provide run time in days.
2. Output is given per flow cell. HiSeq 2500, HiSeq 4000, and NovoSeq 6000 are able to run one or two flow cells simultaneously.
3. NextSeq 500 is suitable for batches of 10-20 samples where rapid turn-around is required.
4. Service on NovoSeq 6000 is not yet available at ORB. Run time, and read output are given for the S2 kit configuration. Kit configurations S1, S3, and S4 are expected to yield 1.6, 6.6, and 10 billion reads per flow cell, respectively.