As a step towards simplifying and reducing the cost of haplotype resolved assembly, we describe new methods for accurately phasing nanopore data with the Shasta genome assembler and a modular tool for extending phasing to the chromosome scale called GFAse. We test using new variants of Oxford Nanopore Technologies' (ONT) PromethION sequencing, including those using proximity ligation and show that newer, higher accuracy ONT reads substantially improve assembly quality.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9980101PMC
http://dx.doi.org/10.1101/2023.02.21.529152DOI Listing

Publication Analysis

Top Keywords

phased nanopore
4
nanopore assembly
4
assembly shasta
4
shasta modular
4
modular graph
4
graph phasing
4
phasing gfase
4
gfase step
4
step simplifying
4
simplifying reducing
4

Similar Publications

Structural variants (SVs) drive gene expression in the human brain and are causative of many neurological conditions. However, most existing genetic studies have been based on short-read sequencing methods, which capture fewer than half of the SVs present in any one individual. Long-read sequencing (LRS) enhances our ability to detect disease-associated and functionally relevant structural variants (SVs); however, its application in large-scale genomic studies has been limited by challenges in sample preparation and high costs.

View Article and Find Full Text PDF

Somatic mutations in individual cells lead to genomic mosaicism, contributing to the intricate regulatory landscape of genetic disorders and cancers. To evaluate and refine the detection of somatic mosaicism across different technologies with personalized donor-specific assembly (DSA), we obtained tissue from the dorsolateral prefrontal cortex (DLPFC) of a post-mortem neurotypical 31-year-old individual. We sequenced bulk DLPFC tissue using Oxford Nanopore Technologies (∼60X), NovaSeq (∼30X), and linked-read sequencing (∼28X).

View Article and Find Full Text PDF

Advanced phasing techniques in congenital skin diseases.

J Dermatol

December 2024

Department of Dermatology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, Japan.

Phasing, the process of determining which alleles at different loci on homologous chromosomes belong together on the same chromosome, is crucial in the diagnosis and management of autosomal recessive diseases. Advances in long-read sequencing technologies have significantly enhanced our ability to accurately determine haplotypes. This review discusses the application of low-coverage long-read sequencing, nanopore Cas9-guided long-read sequencing, and adaptive sampling in phasing, highlighting their utility in complex clinical scenarios.

View Article and Find Full Text PDF
Article Synopsis
  • Long-read technologies from PacBio and ONT have revolutionized genomics research, but there are ongoing challenges in representing genetic diversity and assembling comprehensive pangenomes.
  • The study investigates the necessary data types and volumes for effective de novo genome assembly in pangenome projects, comparing the performance of ONT's Duplex and PacBio HiFi datasets.
  • Results indicate that achieving high-quality phased genomes requires significant amounts of long reads and various supplemental data, with PacBio HiFi showing better phasing accuracy and ONT Duplex producing more complete contigs.
View Article and Find Full Text PDF

Phasing nanopore genome assembly by integrating heterozygous variations and Hi-C data.

Bioinformatics

November 2024

School of Computer Science and Engineering, Central South University, Changsha, Hunan 410083, China.

Motivation: Haplotype-resolved genome assemblies serve as vital resources in various research domains, including genomics, medicine, and pangenomics. Algorithms employing Hi-C data to generate haplotype-resolved assemblies are particularly advantageous due to its ready availability. Existing methods primarily depend on mapping quality to filter out uninformative Hi-C alignments which may be susceptible to sequencing errors.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!