See More,
Discover More

Long-Read Sequencing Delivers
More Data, More Insights

See More,
Discover More

Long-Read Sequencing Delivers More Data, More Insights

Limitless Possibilities

Why long-read sequencing?

Long-read sequencing provides enhanced structural variant detection, true variant phasing, full-length transcript analysis, and high-resolution epigenetic characterization. Generate high-quality data with reads from a few thousand bases to up to 100,000 bases in a single read. Long-read sequencing reveals regions of the genome that are difficult or impossible to sequence using short reads. Plus, long-read sequencing contains built-in DNA modification information, such as methylation – no need for additional sequencing. See what you’ve been unable to see in the past with long-read sequencing.

Why Long-Read Sequencing?

Long-read sequencing provides enhanced structural variant detection, true variant phasing, full-length transcript analysis, and high-resolution epigenetic characterization. Generate high-quality data with reads from a few thousand bases to up to 100,000 bases in a single read. Long-read sequencing reveals regions of the genome that are difficult or impossible to sequence using short reads. Plus, long-read sequencing contains built-in DNA modification information, such as methylation – no need for additional sequencing. See what you’ve been unable to see in the past with long-read sequencing.

Starting on the right foot

The first step in harnessing the power of long-read sequencing is obtaining intact and clean high molecular weight (HMW) DNA and full-length, intact RNA for your studies.

Next Level Genomics provides DNA and RNA extraction services geared toward long-read sequencing, allowing you to take full advantage of the power of LRS – quicker and easier. Our experts have worked on a wide range of sample types including blood, animal tissues, cells, environmental samples and more.

Let Next Level Genomics get you started on the right foot with our HMW DNA and Ultrapure RNA extraction services.

Starting on the right foot

The first step in harnessing the power of long-read sequencing is obtaining intact and clean high molecular weight (HMW) DNA and full-length, intact RNA for your studies.

Next Level Genomics provides DNA and RNA extraction services geared toward long-read sequencing, allowing you to take full advantage of the power of LRS – quicker and easier. Our experts have worked on a wide range of sample types including blood, animal tissues, cells, environmental samples and more.

Let Next Level Genomics get you started on the right foot with our HMW DNA and Ultrapure RNA extraction services.

See More Sequence

Long-Read Sequencing

Long-read sequencing opens up a whole new world of genomics data:

True complete genomes with previously un-attainable regions (dark regions)

Highest resolution and microbiome analysis with whole rRNA/ITS genes or shotgun metagenomics

Complete iso-form information from bulk or single-cell RNA

Structural variation, copy number repeats and previously unseen variants

Native DNA modifications – methylation built-in – for epigenetics and expression analysis

See More Sequence

Long-Read Sequencing

Long-read sequencing opens up a whole new world of genomics data:

True complete genomes with previously un-attainable regions (dark regions)​

Highest resolution and microbiome analysis with whole rRNA/ITS genes or shotgun metagenomics​

Complete iso-form information from bulk or single-cell RNA​

Structural variation, copy number repeats and previously unseen variants​

Native DNA modifications – methylation built-in – for epigenetics and expression analysis​

DNA Sequencing

Long-read sequencing has revolutionized DNA sequencing, offering ease, affordability, and productivity. With the ability to sequence from a few kilobases to over 100 kilobases per read, de novo genome assembly has become simpler. This advancement enabled the completion of the first truly comprehensive human genome, the Telomere-to-Telomere assembly (T2T), in 2022 

With long-read sequencing technologies, DNA can be sequenced for

Complete variant analysis of human and other reference genomes (SNPs, SVs, CNVs and more)

Metagenomic and microbiome population analysis using either complete 16 rRNA gene sequencing or shotgun

Antimicrobial resistance research based on complete sequence of antibiotic resistance gene carrying plasmids

De novo assembly of new genomes

Epigenetic analysis from built-in methylation and other modification detection

DNA Sequencing

DNA Sequencing

Sequencing DNA using long-read sequencing technologies has never been easier, more affordable and more productive. The ability to sequence a few kilobases per read, up to over 100 kilobases in a read, has made do novo genome assembly a much easier task and was the sole reason the first truly complete human genome – the Telomere-to-Telomere assembly (T2T) – was able to be completed in 2022 

With long-read sequencing technologies, DNA can be sequenced for

Complete variant analysis of human and other reference genomes (SNPs, SVs, CNVs and more)

Metagenomic and microbiome population analysis using either complete 16 rRNA gene sequencing or shotgun

Antimicrobial resistance research based on complete sequence of antibiotic resistance gene carrying plasmids

De novo assembly of new genomes

Epigenetic analysis from built-in methylation and other modification detection

Complete variant analysis of human and other reference genomes (SNPs, SVs, CNVs and more)

Metagenomic and microbiome population analysis using either complete 16 rRNA gene sequencing or shotgun

Antimicrobial resistance research based on complete sequence of antibiotic resistance gene carrying plasmids

De novo assembly of new genomes

Epigenetic analysis from built-in methylation and other modification detection

In the past, understanding gene expression through the use of RNA sequencing – aka transcriptomics – has had to rely on partial, fragmented data from short read sequencing. The use of long-read sequencing has revolutionized the field of transcriptomics by allowing the complete sequencing of full length mRNA transcripts in a single read – which allows for a complete understanding of the structure of the mRNA, including all alternatively spliced variants, or isoforms. 

RNA Sequencing

RNA Sequencing

In the past, understanding gene expression through the use of RNA sequencing – aka transcriptomics – has had to rely on partial, fragmented data from short read sequencing. The use of long-read sequencing has revolutionized the field of transcriptomics by allowing the complete sequencing of full length mRNA transcripts in a single read – which allows for a complete understanding of the structure of the mRNA, including all alternatively spliced variants, or isoforms. 

With long-read sequencing technologies, RNA can be sequenced for

Combined with single-cell RNA indexing, the use of long-read RNA sequencing has uncovered a myriad of novel isoforms and isoform expression patterns not capable of detection with short read sequencing. 

Complete Iso-form sequencing from bulk RNA

Single-cell isoform sequencing

Complete Iso-form sequencing from bulk RNA

Single-cell isoform sequencing

Platforms

Unlock the power of long-read sequencing with intact, high molecular weight (HMW) DNA and RNA. Next Level Genomics offers HMW DNA and Ultrapure RNA extraction services, ensuring a strong start for your long-read sequencing journey. Trust our experts to handle a variety of sample types, from blood to environmental samples, allowing you to harness the full potential of long-read sequencing quickly and effortlessly

PacBio

Both PacBio systems generate some of the highest quality reads – HiFi Reads – through the generation of Circular Consensus Sequences (CCS) – with read lengths up to 40kB

Revio

Sequel IIe

Higher density
25M SMRT Cell – up to 90Gb data
8M SMRT Cell – up to 30Gb data
Independent stages
Up to 4 cells – run in-parallel
Adaptive loading – maximize output
Capability
Over 1100 WGS per year
Amplicons to WGS

Platforms

PacBio

PacBio

Both PacBio systems generate some of the highest quality reads – HiFi Reads – through the generation of Circular Consensus Sequences (CCS) – with read lengths up to 40kB

Sequel IIe

Revio
Machine Type
Sequel-Ile
25M SMRT Cell – up to 90Gb data
Technical Details
8M SMRT Cell – up to 30Gb data
Up to 4 cells – run in-parallel
Independent Stages
Adaptive loading – maximize output
Over 1100 WGS per year
Capability
Amplicons to WGS

PromethION-24

Oxford Nanopore Technologies

Oxford Nanopore Technologies

Oxford Nanopore Technologies  sequences directly from native DNA or native RNA and detects several base modifications for built-in epigenetic analysis and complete RNA iso-form information

Read lengths from 20bp to ultra-long reads (ULR) possible (N50 > 50kb)

Utilized to generate the fastest human WGS on record (*Stanford University, Dr.  Euan Ashley, 5 hrs)

Machine Type
PromethION-24
Technical Details
Real-time data – terabase scale
Independent Stages
Up to 24 flow cells – like having 24 sequencers
Capability
Over 2400 flow cells per year

Auxillary Long-Read Services

DNA & RNA Extraction

We are experts in extraction of high molecular weight (HMW) DNA and intact RNA

Nucleic Acid Analysis

Determine the purity and quantity of your samples using specialized equipment for long-read analysis

Microbiome 165 PCR

We can amplify barcoded complete, full-length 165 rRNA gene from your microbial samples ready for long-read sequencing

Applications of Long-Read Sequencing

Complete Variant Analysis

  • Sequence complex genomic regions, including repetitive sequences, structural variations, and challenging genomic area

  • Detect large and complex structural variations (SVs) in the genome. SVs, such as deletions, insertions, duplications, inversions, and translocation

  • Identify compound heterozygotes, haplotypes, and allele-specific expression, which can contribute to improved drug target identification and patient stratification

Rare Disease

  • Discover causative variants of rare, undiagnosed conditions

  • Find novel variants in disease or population cohorts

  • Perform disease diagnosis and genetic stratification in disease research or clinical trials

  • Contribute to translational medicine and discoveries of disease and drug targets, which makes it possible to develop new
    treatment strategies

Epigenetics

  • Methylation information built-in – no additional sequencing needed

  • Simplified haplotype phasing of methylated bases using long-reads

  • Capability to interrogate imprinting disorders and methylation abnormalities associated with tandem repeats

  • Microbial epigenetics – Detect genome-wide m6A and m4C R-M system motifs

Dark Regions

  • Twist Alliance Dark Genes panel for PacBio, a comprehensive 22 Mb panel of 389 medically-relevant dark genes, including CYP2D6, GBA, SMN1/2, and PMS2

  • Adaptive Sampling with Oxford Nanopore allows for region-specific targeted sequencing

  • Find genes missed by short-read NGS that might typically require several platforms to adequately capture, allowing for assay consolidation and cost-effectiveness

     

Pharmacogenomics

  • Comprehensive 49-gene PGx panel from Twist / Pac-Bio Community Alliance for long-read sequencing that includes all 20 current genes with CPIC guidelines, as well as FDA PGx genes, and genes research interest

  • Targeted haplotyping in pharmacogenomics using adaptive sampling using Oxford Nanopore from DNA or RNA

Targeted Gene Panels

  • Sequence only the genomic regions you care about — easily and cost-effectively at scale

  • Adaptive sampling allows for targeted sequencing without capture – both DNA and RNA adaptive sampling

  • Hybrid capture and amplicon workflows are also available to suit your needs

  • Pre-designed panels or custom panels available

  • Span SVs end-to-end enabling unprecedented resolution of even highly complex variants — in any genomic context

De Novo Genomes

  • Build reference-quality, haplotype-resolved genomes and pangenomes to better understand individuals, populations, and species

  • Impute traits of interest to SNPs, structural variants, and complex genotypes

  • Capture genomic variants on a genome-wide scale for outbreds, inbreds, and populations

Pan-genomes

  • Long-read sequencing has revolutionized the study of pan-genomes beyond microbial genomes

  • Capture the entire set of genes within a species, consisting of a core genome — containing sequences shared between all individuals of the species — and the ‘dispensable’ genome

  • Pan genome analysis allows for the discovery of variants that are missing from the reference genome but may be associated with specific phenotypes

Direct, Native RNAseq

  • Nanopore direct single molecule RNA sequencing, is highly parallel, real-time, single-molecule method without prior conversion of RNA to cDNA

  • Low input amounts and streamlined workflows enable highly sensitive gene expression analysis

  • Full-length transcripts of all iso-forms reads delivered by nanopore sequencing allow identification of splice variants and gene fusions

Isoform Sequencing

  • Discover alternative splicing events, including alternative start sites, end sites, intron retention, and exon-skipping events

  • Find novel gene fusions and identify allele-specific isoforms

  • Detect differentially expressed isoforms down to single-cell resolution

Single-cell RNAseq

  • From input single-cell cDNA, utilizing novel concatenation methods, generate a 16-fold throughput increase compared to regular single-cell Iso-Seq libraries with new kits from PacBio

  • Available on both PacBio and Oxford Nanopore platforms

  • Compatible with cDNA generated using the 10x Chromium Next GEM Single Cell 3’ kit (v3.1) and Single Cell 5’ kit (v2) and is intended for use on a 3,000 to 10,000 cell library with 15-75 ng of cDNA as input

Targeted RNAseq

  • Utilize the adaptive sampling function of Oxford Nanopore sequencing to selectively deplete and enrich RNAs of interest without biochemical manipulation before sequencing

  • Identify previously unknown transcripts and isoforms, including those which are antisense to existing transcripts

  • Depleting unwanted transcripts can enrich RNAs of interest

Whole Microbial Genomes

  • Generate the highest quality, closed reference genomes

  • Identify ever-evolving genes associated with toxicity, virulence, and antimicrobial resistance

  • Resolve strains, serotypes, and plasmids to track pathogen outbreaks in humans, plants, and animals, through food systems, hospitals, and communities

  • Comprehensively characterize microbes to facilitate scientific breakthroughs

Full Length 16S rRNA

  • Overcome misclassification at the species-level using full-length 16S rRNA sequencing with long reads to resolve similarity in each 16S variable region

  • The Kinnex 16S rRNA kits from PacBio increase throughput, increasing the resolution of microbial analysis

  • Massive multiplexing capabilities on both PacBio and Oxford Nanopore platforms

Shotgun Metagenomics

  • Identify 6-8 full-length genes with efficient, cost-effective metagenomic profiling

  • Generate hundreds of high-quality (HQ) metagenome assembled genomes (MAGs), many of which are single contig, circular MAGs

  • Leverage epigenomic data to associate contigs and plasmids from closely related strains

Microbiomes

  • Determine community composition at the species or strain-level with competitively priced full-length 16S sequencing

  • Achieve the taxonomic and functional resolution needed to generate testable hypotheses about a microbiota’s impact on health, disease, the environment and more

  • Resolve structural variants in the microbiome and expand the detection scope of genetic variations, enabling profiling of strain-level variation

Auxillary Long-Read Services

DNA & RNA Extraction

We are experts in extraction of high molecular weight (HMW) DNA and intact RNA

Nucleic Acid Analysis

Determine the purity and quantity of your samples using specialized equipment for long-read analysis

Microbiome 165 PCR

We can amplify barcoded complete, full-length 165 rRNA gene from your microbial samples ready for long-read sequencing

Frequently Asked Questions

How do I start a long-read sequencing project?

The best place to start is to contact our NGS specialists to discuss your project goals and needs. Even prior to beginning a project, we can advise on ways to make the best use of long-read sequencing – from extraction method to multiplexing to sequencing strategies – we are there to help you get the most out of your long-read project in the most efficient and economical manner. We can pass along publications, tech notes and other materials related to your project to demonstrate the power of long-read sequencing and benefits you’ll obtain.

For samples within Singapore, Next Level Genomics can arrange for sample collection from your lab at your convenience. Alternatively, samples can be dropped off at our labs in Biopolis. We are happy to discuss your project with you and what works the best for getting your samples to our lab in the best manner. Samples from outside Singapore can be shipped directly to our labs based on your needs.

Each project is unique – sample type, sample number and output requirements. For whole genome sequencing of DNA, libraries can often be prepared within 1-2 weeks of quality clearance of the sample. Long read sequencing generally takes 36-40 hours. With the Revio system, multiple samples can run in parallel, shortening the run time for a project. Spatial slides can be processed in 1-2 weeks and spatial imaging or profiling can last from hours to days, depending on the project. Contact our lab specialists to discuss your project details to learn more about how we can serve you best and most efficiently.

Next Level Genomics provides bioinformatics and data analysis services utilizing a combination of tools provided by the equipment manufacturers such as Smart Link from PacBio and AtoMx from Nanostring – as well as custom bioinformatics to suite your needs. Data can be delivered to you via cloud, FTP or physical hard drive. Speak with our specialists to learn how Next Level Genomics can provide complete end-to-end service from sample extraction and handling to data generation and full analysis.

Yes, Next Level Genomics has all the equipment and experience in DNA and RNA extraction to take your primary samples and perform complete end-to-end processing. As long-read sequencing experts, we have experience in isolation of high-molecular-weight (HMW) DNA that is perfect for both PacBio and Oxford Nanopore sequencing. Speak to our specialists to learn how we can assist in getting your project off on the right foot with optimum extraction.

Read length can vary based on your project needs. Microbial genomes and targeted approaches can focus on reads between 5kb-10kb, while whole genome analysis may need 15kb-20kb. With Nanopore sequencing, ultra-long reads of 50kb or larger are possible. The amount of data generated can be customized to your project based utilizing multiplexing and barcode to control the number of samples on a flowcell and data generated. Human genomes from 25X-30X are possible on a single flow cell with both systems Next Level Genomics uses.

Yes, with UMIs and barcoded adapters, samples can be individually tagged and multiplexed to customized the amount of data generated per sample – this can often account for significant savings on costs of long-read sequencing. In addition, by submitted multiple samples for processing all at the same time, savings can be had by batching the libraries during the quality control (QC) steps.

With both long-read platforms – PacBio and Oxford Nanopore – DNA modifications are including the sequencing results – built-in. No additional treatments or sequencing is needed to obtain epigenetic information from your samples. Discuss your needs with our specialists to learn how built-in modification information can benefit your project.

Talk with an Expert

Unlock the potential of your genomics projects with personalized guidance. Connect with our seasoned experts at Next Level Genomics to discuss your goals, explore possibilities, and pave the way for groundbreaking discoveries. Fill out the form to initiate a discussion and propel your research forward.