scholarly journals Investigational Assay for Haplotype Phasing of the Huntingtin Gene

2020 ◽  
Vol 19 ◽  
pp. 162-173
Author(s):  
Nenad Svrzikapa ◽  
Kenneth A. Longo ◽  
Nripesh Prasad ◽  
Ramakrishna Boyanapalli ◽  
Jeffrey M. Brown ◽  
...  
Keyword(s):  
Haplotype Phasing

Haplotype phasing of whole human genomes using bead-based barcode partitioning in a single tube

2017 ◽  
Vol 35 (9) ◽  
pp. 852-857 ◽  
Author(s):  
Fan Zhang ◽  
Lena Christiansen ◽  
Jerushah Thomas ◽  
Dmitry Pokholok ◽  
Ros Jackson ◽  
...  
Keyword(s):  
Single Tube ◽  
Haplotype Phasing ◽  
Human Genomes

scholarly journals stLFRsv: A Germline Structural Variant Analysis Pipeline Using Co-barcoded Reads

2021 ◽  
Vol 12 ◽  
Author(s):  
Junfu Guo ◽  
Chang Shi ◽  
Xi Chen ◽  
Ou Wang ◽  
Ping Liu ◽  
...  
Keyword(s):  
Structural Variation ◽  
Signal To Noise Ratio ◽  
Recall Rate ◽  
Structural Variants ◽  
Analysis Pipeline ◽  
Haplotype Phasing ◽  
Base Level ◽  
Long Reads ◽  
Variant Analysis ◽  
Complex Structural

Co-barcoded reads originating from long DNA fragments (mean length >30 kbp) maintain both single base level accuracy and long-range genomic information. We propose a pipeline, stLFRsv, to detect structural variation using co-barcoded reads. stLFRsv identifies abnormal large gaps between co-barcoded reads to detect potential breakpoints and reconstruct complex structural variants (SVs). Haplotype phasing by co-barcoded reads increases the signal to noise ratio, and barcode sharing profiles are used to filter out false positives. We integrate the short read SV caller smoove for smaller variants with stLFRsv. The integrated pipeline was evaluated on the well-characterized genome HG002/NA24385, and 74.5% precision and a 22.4% recall rate were obtained for deletions. stLFRsv revealed some large variants not included in the benchmark set that were verified by long reads or assembly. For the HG001/NA12878 genome, stLFRsv also achieved the best performance for both resource usage and the detection of large variants. Our work indicates that co-barcoded read technology has the potential to improve genome completeness.

scholarly journals LRSim: a Linked Reads Simulator generating insights for better genome partitioning

2017 ◽  
Author(s):  
Ruibang Luo ◽  
Fritz J. Sedlazeck ◽  
Charlotte A. Darby ◽  
Stephen M. Kelly ◽  
Michael C. Schatz
Keyword(s):  
Genome Assembly ◽  
Fragment Length ◽  
De Novo ◽  
Library Preparation ◽  
Haplotype Phasing ◽  
Multiple Datasets ◽  
Long Read ◽  
Fine Control ◽  
Informatics Tools ◽  
Sequencing Process

AbstractMotivationLinked reads are a form of DNA sequencing commercialized by 10X Genomics that uses highly multiplexed barcoding within microdroplets to tag short reads to progenitor molecules. The linked reads, spanning tens to hundreds of kilobases, offer an alternative to long-read sequencing for de novo assembly, haplotype phasing and other applications. However, there is no available simulator, making it difficult to measure their capability or develop new informatics tools.ResultsOur analysis of 13 real linked read datasets revealed their characteristics of barcodes, molecules and partitions. Based on this, we introduce LRSim that simulates linked reads by emulating the library preparation and sequencing process with fine control of 1) the number of simulated variants; 2) the linked-read characteristics; and 3) the Illumina reads profile. We conclude from the phasing and genome assembly of multiple datasets, recommendations on coverage, fragment length, and partitioning when sequencing human and non-human genome.AvailabilityLRSIM is under MIT license and is freely available at https://github.com/aquaskyline/[email protected]

scholarly journals Targeted isolation of cloned genomic regions by recombineering for haplotype phasing and isogenic targeting

2011 ◽  
Vol 39 (20) ◽  
pp. e137-e137 ◽  
Author(s):  
Marta Nedelkova ◽  
Marcello Maresca ◽  
Jun Fu ◽  
Maria Rostovskaya ◽  
Ramu Chenna ◽  
...  
Keyword(s):  
Haplotype Phasing ◽  
Genomic Regions

scholarly journals Universal Haplotype-Based Noninvasive Prenatal Testing for Single Gene Diseases

2017 ◽  
Vol 63 (2) ◽  
pp. 513-524 ◽  
Author(s):  
Winnie W I Hui ◽  
Peiyong Jiang ◽  
Yu K Tong ◽  
Wing-Shan Lee ◽  
Yvonne K Y Cheng ◽  
...  
Keyword(s):  
Single Gene ◽  
Family Members ◽  
Prenatal Testing ◽  
Maternal Plasma ◽  
Nucleotide Polymorphisms ◽  
Plasma Dna ◽  
Sequencing Technology ◽  
Noninvasive Prenatal Testing ◽  
Haplotype Phasing ◽  
Mutational Status

Abstract BACKGROUND Researchers have developed approaches for the noninvasive prenatal testing of single gene diseases. One approach that allows for the noninvasive assessment of both maternally and paternally inherited mutations involves the analysis of single nucleotide polymorphisms (SNPs) in maternal plasma DNA with reference to parental haplotype information. In the past, parental haplotypes were resolved by complex experimental methods or inferential approaches, such as through the analysis of DNA from other affected family members. Recently, microfluidics-based linked-read sequencing technology has become available and allows the direct haplotype phasing of the whole genome rapidly. We explored the feasibility of applying this direct haplotyping technology in noninvasive prenatal testing. METHODS We first resolved the haplotypes of parental genomes with the use of linked-read sequencing technology. Then, we identified SNPs within and flanking the genes of interest in maternal plasma DNA by targeted sequencing. Finally, we applied relative haplotype dosage analysis to deduce the mutation inheritance status of the fetus. RESULTS Haplotype phasing and relative haplotype dosage analysis of 12 out of 13 families were successfully achieved. The mutational status of these 12 fetuses was correctly classified. CONCLUSIONS High-throughput linked-read sequencing followed by maternal plasma-based relative haplotype dosage analysis represents a streamlined approach for noninvasive prenatal testing of inherited single gene diseases. The approach bypasses the need for mutation-specific assays and is not dependent on the availability of DNA from other affected family members. Thus, the approach is universally applicable to pregnancies at risk for the inheritance of a single gene disease.

scholarly journals Noninvasive Prenatal Diagnosis for Duchenne Muscular Dystrophy Based on the Direct Haplotype Phasing

2019 ◽  
Author(s):  
Min Chen ◽  
Chao Chen ◽  
Xiaoyan Huang ◽  
Jun Sun ◽  
Lu Jiang ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Chorionic Villus ◽  
Chorionic Villus Sampling ◽  
Singleton Pregnancy ◽  
High Molecular Weight Dna ◽  
Haplotype Phasing ◽  
Noninvasive Prenatal Diagnosis ◽  
Mutational Status

AbstractObjectiveWe aimed to investigate the validity of noninvasive prenatal diagnosis (NIPD) based on direct haplotype phasing without the proband and its feasibility for clinical application in the case of Duchenne Muscular Dystrophy (DMD).MethodsThirteen singleton-pregnancy families affected by DMD were recruited. Firstly, we resolved maternal haplotypes for each family by performing targeted linked-read sequencing of their high molecular weight DNA, respectively. Then, we identified SNPs of the DMD gene in all carrier mothers and inferred the DMD gene mutation status of all fetuses. Finally, the fetal genotypes were further validated by using chorionic villus sampling.ResultsThe method of directly resolving maternal haplotype through targeted linked-read sequencing was smoothly performed in all participated families. The predicted mutational status of 13 fetuses was correct, which had been confirmed by invasive prenatal diagnosis.ConclusionDirect haplotyping of NIPD based on linked-read sequencing for DMD is accurate.

scholarly journals Ultra-low input single tube linked-read library method enables short-read NGS systems to generate highly accurate and economical long-range sequencing information for de novo genome assembly and haplotype phasing

2019 ◽  
Author(s):  
Zhoutao Chen ◽  
Long Pham ◽  
Tsai-Chin Wu ◽  
Guoya Mo ◽  
Yu Xia ◽  
...  
Keyword(s):  
Long Range ◽  
De Novo ◽  
Low Cost ◽  
Cost Effective ◽  
De Novo Genome Assembly ◽  
Short Read ◽  
Single Tube ◽  
Haplotype Phasing ◽  
A Genome ◽  
Long Read

AbstractLong-range sequencing information is required for haplotype phasing, de novo assembly and structural variation detection. Current long-read sequencing technologies can provide valuable long-range information but at a high cost with low accuracy and high DNA input requirement. We have developed a single-tube Transposase Enzyme Linked Long-read Sequencing (TELL-Seq™) technology, which enables a low-cost, high-accuracy and high-throughput short-read next generation sequencer to routinely generate over 100 Kb long-range sequencing information with as little as 0.1 ng input material. In a PCR tube, millions of clonally barcoded beads are used to uniquely barcode long DNA molecules in an open bulk reaction without dilution and compartmentation. The barcode linked reads are used to successfully assemble genomes ranging from microbes to human. These linked-reads also generate mega-base-long phased blocks and provide a cost-effective tool for detecting structural variants in a genome, which are important to identify compound heterozygosity in recessive Mendelian diseases and discover genetic drivers and diagnostic biomarkers in cancers.

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