whole genome sequencing
About
Nanopore sequencing is a unique, scalable technology that enables direct, real-time analysis of long DNA or RNA fragments. It works by monitoring changes to an electrical current as nucleic acids are passed through a protein nanopore. The resulting signal is decoded to provide the specific DNA or RNA sequence.
About
Nanopore sequencing is a unique, scalable technology that enables direct, real-time analysis of long DNA or RNA fragments. It works by monitoring changes to an electrical current as nucleic acids are passed through a protein nanopore. The resulting signal is decoded to provide the specific DNA or RNA sequence.
Oxford Nanopore Technologies (ONT) enables real-time, long-read sequencing of native DNA and RNA, eliminating PCR bias. The technology feeds single-stranded DNA or RNA molecules through a protein nanopore to detects changes in electrical current. This preserves strand methylation and allows direct detection of DNA/RNA modifications without the need for chemical conversion.
Long reads from ONT outperform traditional short-read sequencing in resolving repeat regions and structural variants. These reads of 10–100 kb or longer enable detection of SNVs, SVs, indels, CNVs, STRs, and genome-wide methylation patterns, providing a more complete view of genetic variation, including phasing.
Samples are sequenced with the PromethION P24, followed by filtering, assembly, annotation, and quality control using ONT’s EPI2ME software and a data analysis pipelines developed by Poochon Scientific.
+ W101: Whole Genome Sequencing
Nanopore Long-Read Whole Genome Sequencing
Turnaround Time: 7 to 15 business days
Service Description:
Construction of an amplification-free long-read sequencing library using the newest ONT v14 library prep chemistry and data acquisition by ONT PromethION P24 (bases up to 190 GB, coverage up to 60X, N50 read length from 10 Kb to > 100 Kb)
Deliverables:
Raw data files
1) Fasta.gz – a compressed file of all the raw ONT sequencing reads
2) BAM (upon request)
Analysis Report Files
1) Reports (html) – Alignment/SNV/STR/SV/CNV
2) .fasta – polished consensus sequence of the genome (may contain multiple contigs)
3) VCF files
4) TSV files
5) Other QC files
Suitable Sample Type:
≥ 5 µg of genomic DNA from any species: OD260/OD280 = ~1.8; OD260/OD230 = ~2.0-2.2; average size ~50 Kb; concentration ≥ 200 ng/µl; buffer: 10mM Tris pH 8.0, EB, or low TE (< 0.1 mM EDTA)
Or, cell pellet samples from any species (≥5 million cells per sample)
Sample Preparation Guidelines
Sample Type
Genomic DNA (≥ 5 µg) or cell pellet (≥5 million cells ) from any species
Size
Average size ~50 Kb
OD Ratio
1.8 ≤ OD 260/280 ≤ 1.95
2.0 ≤ OD 260/230 ≤ 2.2
Concentration
≥ 200 ng/µl
Buffer
ddH2O, EB, or low TE (< 0.1 mM EDTA)
Advantages of Whole Genome Sequencing by Nanopore Technology
- Generation of ultra-long reads (50–100 kb+, up to 4 Mb)
- Comprehensive analysis of SNVs, SVs, indels, CNVs, STRs, and methylation
- Detection of complex structural variation and repetitive regions
- Direct methylation analysis from native DNA
