Nucleome Informatics is the only company in India to offer sequencing services on the PacBio Sequel II system, utilizing the highly accurate HiFi sequencing technology. We provide whole-genome sequencing services for Microbe, Plant, and Animal Genomes.
Sample Requirement:
HMW gDNA and Library QC:
HMW gDNA Shearing and Size Selection:
Library Insert Size:
Sequencing on Sequel II Platform:
PacBio HiFi Coverage:
Data Analysis:
We use state-of-the-art tools to assemble genomes, building contig-level assemblies with PacBio HiFi data. Genomes are scaffolded with Optical Map and HiC data after multiple rounds of polishing, gap filling, and curation. Assembly quality is assessed using metrics such as Contig N50, Scaffold N50, number of gaps, base accuracy, BUSCO score, RNA-Seq mappability, phase block resolution, and chromosomal-level continuity. Our services include:
For any queries or more information about our services, don’t hesitate to contact our sales team at sales @ nucleomeinfo.com.
Unlock the full potential of genomic insights with Nucleome’s cutting-edge HiFi Sequencing service, designed to provide highly accurate and comprehensive structural variant (SV) detection in human genomes. Leveraging PacBio’s revolutionary HiFi reads, our service delivers unparalleled resolution for complex genetic analysis, making it an ideal choice for researchers studying rare and inherited diseases.
HiFi reads combine long-read length with exceptional accuracy (99.9%), empowering researchers to:
Variation between two human genomes, by the number of base pairs impacted, reveals that single-nucleotide variants (SNVs) are the most numerous variant type (~4–5 M/person), followed by indels. Structural variants (SVs) and tandem repeats, while fewer in number, account for more genomic variation between individuals due to their size (>50 bp). This makes them critical for understanding genetic diversity and disease associations.
HiFi sequencing stands out by offering accurate small variant calls alongside significantly more structural variant detection compared to other technologies. This comprehensive resolution enables researchers to:
HiFi sequencing is transforming how researchers approach rare disease diagnostics by:
The extraordinary accuracy and long read lengths of PacBio® HiFi sequencing overcome the challenges of detecting variation in cancer genomes faced by other technologies.
High-quality HiFi reads detect a wide range of cancer-specific somatic variation, including single nucleotide variants (SNVs), structural variants (SVs), insertions and deletions (indels), copy number variations (CNVs), and methylation in a single assay. With improved somatic variant calling accuracy, HiFi sequencing requires less sequencing relative to other technologies, making it an efficient solution for cancer research.
As an experienced long-read sequencing technology user, Nucleome Informatics is committed to providing world-class sequencing solutions. Our HiFi Sequencing service for SV discovery and somatic cancer variant detection empowers researchers to uncover the genetic basis of diseases with unmatched accuracy and efficiency.
Contact us today to explore how HiFi sequencing can revolutionize your research.
Nucleome provides full-length transcriptome sequencing i.e ISO Sequencing on Pacbio Sequel II platform. Nucleome Informatics is the only service provider of ISO Sequencing on Sequel II in India. The PacBio Iso-Seq method is an end-to-end workflow for sequencing and analyzing full-length transcript isoforms. Our Iso-Seq analysis enables you to detect novel transcripts and genes, identify fusion genes, annotate isoforms and alternative splicing events.
Generate full-length transcript sequences up to ~15kb
ISO Seq Service WorkFLow:
1. Convert RNA into cDNA
2. cDNA SMRTbell library preparation
3. Sequence on the Sequel II System
4. Generate circular consensus sequences (CCS)
5. Discover isoforms de novo with Iso-Seq analysis
ISO Seq Applications:
Whole-genome Annotation
-Typically whole-transcriptome, non- quantitative
-Often included in de novo genome assembly projects
-Single tissue to several tissues
-Generates reference transcriptome for downstream RNA-seq studies
Gene-level Isoform Discovery
-Typically targeted, either cDNA amplicons or target capture
-Useful for detecting gene fusions, SNVs, allele-specific expression
-Cost-effectively multiplex many samples per single SMRT Cell
-Relative quantitation possible
Sample Requirement
1 ug of high qulaity total RNA, RIN >7 and Concentration should be more than 25ng per ul
– Has not been exposed to high temperatures (e.g.: > 65°C for 1 hour can cause a detectable decrease in sequence quality) or pH extremes (< 6 or > 9).
– Has an OD260/OD280 ratio between 2.0 and 2.2. – Has an OD260/OD230 ratio between 1.8 and 2.1. – Has a RIN number ≥ 8 (Recommended).
– Has not been exposed to intercalating fluorescent dyes or ultraviolet radiation. SYBR dyes are not RNA damaging, but do avoid ethidium bromide.
– Does not contain denaturants (e.g., guanidinium salts or phenol) or detergents (e.g., SDS or Triton‐X100).
– Does not contain carryover contamination from the original organism/tissue (e.g., heme, humic acid, polyphenols, etc.).
– Note: RNA samples should only be shipped with dry ice.
Contact our team now to receive the quote and place the order.
Nucleome is the only service provider in South Asia that offers Single Molecule, Real-Time (SMRT) Sequencing on PacBio Sequel II that generates the long, accurate, single-molecule reads you need to comprehensively characterize samples with complex variation. With SMRT Sequencing you can:
Highly accurate long reads – HiFi reads – with the single-molecule resolution are ideal for full-length 16S rRNA sequencing, shotgun metagenomic profiling, and metagenome assembly so that you can:
Full-length Metagenome assembly and functional profiling on PacBio Sequel II
HiFi sequencing on the PacBio Sequel II System enables complete microbial community profiling of complex metagenomic samples using whole genome shotgun sequences. With HiFi sequencing, highly accurate long reads overcome the challenges posed by the presence of intergenic and extragenic repeat elements in microbial genomes, thus greatly improving phylogenetic profiling and sequence assembly.
HiFi sequencing can be used even for sample types with limited starting material, allowing for reference quality MAGs, discovery of novel genes without the need for de novo assembly, identification of biochemical pathways, and resolution of taxa below the species level with confidence.
Data analysis:
Output data in standard file formats, (BAM and FASTA/Q) for seamless integration with downstream analysis tools – Perform taxonomic classification and functional gene profiling using QIIME and MEGAN
– Perform gene prediction and discovery using FragGeneScan and Prodigal
Learn more about workflows for these types of complex populations
– Perform metagenomic shotgun assembly directly with HiFi reads using HiCanu
Full length 16s Amplicon Sequencing by Nucleome
Nucleome is the only service provider in South Asia that offers Single Molecule, Real-Time (SMRT) Sequencing on PacBio Sequel II that generates full length 16s amplicon sequencing. The ~1500 bp 16S rRNA gene comprises nine variable regions interspersed throughout the highly conserved 16S sequence. Sequencing the partial gene was originally accomplished by Sanger sequencing. This required cloning genes, generating, and assembling two to three reads per clone, and producing limited sampling depth at high cost and effort. The partial 16s rRNA gene sequencing misses to sequence the entire taxa of the bacterial community.
DIFFERENT SUB-REGIONS SHOWED BIAS IN THE BACTERIAL TAXA THEY WERE ABLE TO IDENTIFY
Full V1–V9 region: the only way to resolve ALL the clades that may be present in a complex population. Order full length 16s Amplicon sequencing to Nucleome and discover Strain-level identification of bacterial communities with the unprecedented accuracy of PacBio HiFi reads.
Nucleome Informatics embraces the power of PacBio Sequel II’s HiFi sequencing technology to shed light on the intricate world of microbial genomes. With this cutting-edge platform, we’ve unlocked the potential to decipher the genetic codes of bacteria, viruses, bacteriophages, and fungal organisms like never before. In this article, we’ll explore how HiFi sequencing on the PacBio Sequel II platform is redefining our understanding of these microscopic life forms. Microbial genome sequencing has revolutionized our understanding of the microbial world. PacBio Sequel II, with its cutting-edge technology, has emerged as a game-changer in the field of microbial genomics. This state-of-the-art platform offers unparalleled advantages for sequencing microbial genomes, enabling researchers to delve deeper into the genetic diversity and functional potential of microorganisms. In this article, we will explore how PacBio Sequel II empowers scientists to unravel the mysteries of microbial genomes.
The Power of Long Reads:
One of the key features that sets PacBio Sequel II apart is its ability to produce long-read sequences. Traditional short-read sequencing technologies often struggle with complex microbial genomes due to their fragmented assembly. PacBio’s Single Molecule, Real-Time (SMRT) sequencing technology generates long reads, making it easier to assemble and analyze microbial genomes, especially those with high GC content or repetitive regions.
High Accuracy and Low Error Rates:
PacBio Sequel II boasts impressive accuracy levels, even in regions that are challenging for other sequencing platforms. With the use of circular consensus sequencing (CCS), it is possible to obtain highly accurate HiFi reads (>99.99% accuracy). This is crucial when studying microbial genomes, as even a single base-pair error can significantly impact downstream analysis, such as functional gene prediction or comparative genomics.
Unraveling Genomic Complexity:
Microbial genomes are incredibly diverse and can contain complex structures, such as plasmids, transposons, and repeat elements. PacBio Sequel II’s long reads and high accuracy help researchers decipher these intricate genomic landscapes with ease. This technology is particularly valuable for investigating microbial populations, identifying genomic variations, and understanding horizontal gene transfer events.
Resolution of Epigenetic Modifications:
In addition to the genetic code, epigenetic modifications play a critical role in microbial function. PacBio Sequel II can identify and characterize DNA modifications, such as methylation patterns, providing insights into gene regulation, adaptation, and virulence mechanisms within microbial populations.
Applications in Microbial Genomics:
The versatility of PacBio Sequel II extends to a wide range of applications in microbial genomics:
Contact us now for receiving the proposal for microbial genome sequencing.