Search results for "DNA sequencing"

Piwi proteins and piRNAs in mammalian oocytes and early embryos: From sample to sequence

2015

AbstractThe role of the Piwi/piRNA pathway during mammalian oogenesis has remained enigmatic thus far, especially since experiments with Piwi knockout mice did not reveal any phenotypic defects in female individuals. This is in striking contrast with results obtained from other species including flies and zebrafish. In mouse oocytes, however, only low levels of piRNAs are found and they are not required for their function. We recently demonstrated dynamic expression of PIWIL1, PIWIL2, and PIWIL3 during mammalian oogenesis and early embryogenesis. In addition, small RNA analysis of human, crab-eating macaque and cattle revealed that piRNAs are also expressed in the female germline and closel…

Sequencing analysis of a 4·1 kb subtelomeric region from yeast chromosome IV identifiesHXT15, a new member of the hexose transporter family

1996

The DNA sequence of a 4·1 kb region of Saccharomyces cerevisiae chromosome IV was determined. This region contains a single open reading frame which codes for a member of the hexose transporter family. This new gene has been named HXT15 according to yeast gene data bases. The sequence has been entered in the EMBL data library under Accession Number X92891.

HLA typing from RNA-Seq sequence reads.

2012

We present a method, seq2HLA, for obtaining an individual's human leukocyte antigen (HLA) class I and II type and expression using standard next generation sequencing RNA-Seq data. RNA-Seq reads are mapped against a reference database of HLA alleles, and HLA type, confidence score and locus-specific expression level are determined. We successfully applied seq2HLA to 50 individuals included in the HapMap project, yielding 100% specificity and 94% sensitivity at a P-value of 0.1 for two-digit HLA types. We determined HLA type and expression for previously un-typed Illumina Body Map tissues and a cohort of Korean patients with lung cancer. Because the algorithm uses standard RNA-Seq reads and …

An approach to determining anthocyanin synthesis enzyme gene expression in an evolutionary context: an example from Erica plukenetii

2019

Abstract Background and Aims Floral colour in angiosperms can be controlled by variations in the expression of the genes of the anthocyanin pathway. Floral colour shifts influence pollinator specificity. Multiple shifts in floral colour occurred in the diversification of the genus Erica (Ericaceae), from plesiomorphic pink to, for example, red or white flowers. Variation in anthocyanin gene expression and its effects on floral colour in the red-, pink- and white-flowered Erica plukenetii species complex was investigated. Methods Next generation sequencing, reverse transcriptase PCR and real-time reverse transcriptase quantitative PCR were used to quantify anthocyanin gene expression. Key Re…

PCR-Typing of the Human HLA-DQα Locus: Population Genetics and Application in Forensic Casework

1991

Multi- and single-locus probes recognizing highly polymorphic DNA sequences throughout the genome ([1–3]; C. Rittner, this volume) have become powerful tools for paternity testing and forensic stain analysis. In forensic casework, however, DNA probe technology can often not be applied, since genomic DNA extracted from stain material exposed to conditions of high temperatures and humidity is degraded. Also, the amount of DNA to be typed may not be sufficient to use DNA probe technology, e.g. if extracted from minute blood or sperm stains, from single hairs or from cell smears on microscope slides.

Molecular biology approaches utilized in preimplantation genetics: real-time PCR, microarrays, next-generation sequencing, karyomapping, and others

2020

Abstract Over the past few decades the development of new molecular technologies has revolutionized diagnosis in the reproductive medicine field, with the evaluation of the two main factors involved in a successful pregnancy: the embryo and the endometrium. The detection of genetically abnormal embryos, as well as the identification of an optimum endometrium using transcriptomics have become a priority in assisted reproductive treatments to increase pregnancy rates. This chapter provides an overview of the molecular techniques currently employed in assisted reproduction for embryo evaluation such as preimplantation genetic testing karyotyping, fluorescence in situ hybridization, polymerase …

Molecular Genetics of Frontonasal Dysplasia

2018

Phylogenomic Analysis Reveals Deep Divergence and Recombination in an Economically Important Grapevine Virus

2015

The evolutionary history of the exclusively grapevine (Vitis spp.) infecting, grapevine leafroll-associated virus 3 (GLRaV-3) has not been studied extensively, partly due to limited available sequence data. In this study we trace the evolutionary history of GLRaV-3, focussing on isolate GH24, a newly discovered variant. GH24 was discovered through the use of next-generation sequencing (NGS) and the whole genome sequence determined and validated with Sanger sequencing. We assembled an alignment of all 13 available whole genomes of GLRaV-3 isolates and all other publicly available GLRaV-3 sequence data. Using multiple recombination detection methods we identified a clear signal for recombinat…

Selection for thermostability can lead to the emergence of mutational robustness in an RNA virus

2010

Mutational robustness has important evolutionary implications, yet the mechanisms leading to its emergence remain poorly understood. One possibility is selection acting on a correlated trait, as for instance thermostability (plastogenetic congruence). Here, we examine the correlation between mutational robustness and thermostability in experimental populations of the RNA bacteriophage Qβ. Thermostable viruses evolved after only six serial passages in the presence of heat shocks, and genome sequencing suggested that thermostability can be conferred by several alternative mutations. To test whether thermostable viruses have increased mutational robustness, we performed additional passages in …

New Vocabulary for Bacterial Communication

2019

Abstract Quorum sensing (QS) is widely accepted as a procedure that bacteria use to converse. However, prevailing thinking places acyl homoserine lactones (AHLs) at the forefront of this communication pathway in Gram‐negative bacteria. With the advent of high‐throughput genomics and the subsequent influx of bacterial genomes, bioinformatics analysis has determined that the genes encoding AHL biosynthesis, originally discovered to be indispensable for QS (LuxI‐like proteins and homologues), are often absent in QS‐capable bacteria. Instead, the sensing protein (LuxR‐like proteins) is present with an apparent inability to produce any outgoing AHL signal. Recently, several signals for these Lux…