WGS Select Long Read

Genome Sequencing Non-Human (Long Read)

Whole Genome Sequencing Non-Human (Long-Read)

Whole Genome Sequencing (WGS) with long-read technology captures DNA in long, continuous stretches, enabling de novo assemblies, the detection of structural variants, highly repetitive regions, and complex rearrangements missed by short-read sequencing. This approach provides more complete genome assemblies, revealing large insertions, deletions, and other structural variations. Additionally, it enhances haplotype phasing, aiding in the distinction between maternal and paternal alleles and allows simultaneous methylation analysis for epigenetic studies.

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Application & Services

Our standard Variant Analysis service includes detection and annotation of germline single nucleotide polymorphism (SNPs), insertions and deletions (InDel).
Additionally, we provide optional services such as, de novo genome assembly, methylation detection, structural variant analysis and copy number variation detection.

Highlights

Choose the instrument that suits your needs best
High quality library preparation for ultra-long reads
Flexible data output for your genomes (depend on the number of genomes loaded per flow cell)
Long term expertise in sequencing (since 2004)

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More details

Specifications

QC of DNA quantity and quality is performed
Library preparation (amplification-free) using v14 library prep chemistry
Sequencing using Oxford Nanopore GridION (flow cell R10.4.1)
Whole flow cells dedicated to your samples
Approx. 6-7 Gb per GridION flow cell (dependent on DNA quality and length). The output is directly dependent on the quantity, quality, and purity of the gDNA sent to us. We do not guarantee any specific output.
Number of samples per flow cell depends on genome size and scientific question:

Theoretical coverage achieved based on genome size and sample number multiplexed per flow cell

			Number of samples
		Genome size (Mb)	1	5	10	30	50
GridION	7 Gb / GridIon FC*	5	1400x	280x	140x	46x	28x
		20	350x	70x	35x	11x
		50	140x	28x	14x
		100	70x	14x

*Expected output of approx. 7GB / GridION flow cell and even read distribution between samples. Long read coverage is calculated as (giga bases (Gb) per flow cell) ÷ (genome size)

Depending on the goals of your studies, we recommend the following coverage for whole genome sequencing:

Germline/frequent variant analysis: 20-50x
De novo assembly: ~100x

Starting material

Sample Type: purified high-molecular weight DNA, with recommended >50% of the DNA above 15 kb in length*
Minimum Amount: 2 µg, recommended 4 µg
Volume: > 50 µL
Concentration: 40 ng/µl using a fluorescence-based method
Purity (OD260/280): 1.8-2.0 with spectrophotometer, e.g. Nanodrop
Resuspension Buffer: Water, EB, or low TE (<0.1 mM EDTA)
Format: barcode labelled 1.5 ml Eppendorf safe-lock tubes
Shipment Method: room temperature

*Please refer to our FAQs for detailed instructions on how to prepare high molecular weight (HMW), RNA- and inhibitor-free double-stranded gDNA.

Deliverables

Raw Nanopore data (.fastq.gz)
Raw Nanopore sequencing data (.pod5) available on request (additional costs)
Additional deliverables for optional bioinformatics, e.g.
- variant analysis package:
  - Alignment file (.bam)
  - SNP and InDel tables including annotated variants and effects (.vcf, .tsv, .bed)
  - Comprehensive data analysis report (.pdf, .html)
- Long-read assembly package:
  - Assembled genome sequence (.fasta)
  - Genomic feature annotation (.gbk, .gff3,. csv, .ffn, .faa)
  - Comprehensive data analysis report (.pdf, .html), including species determination