Gotowa bibliografia na temat „16S amplicon analysis”

Mitochondrial DNA is an important tool for human identification and is used to differentiate between human and animal blood at the crime scene, because in extreme conditions nuclear DNA is severely destroyed while Mitochondrial DNA contains multiple copies (200–2000) per cell and resists harsh and more stable conditions. Seventy-two blood samples were collected from humans (Homo sapiens), sheep (Ovis aries), goats (Capra hircus), and cows (Bos taurus) (18 blood samples for each). All blood samples were withdrawn by a technician and 5 ml were aspirated using an aseptic technique and transferred to EDTA-Na2 tubes. They were mixed well and stored in a refrigerator. The collection took 2 weeks (May 15, 2019–May 30, 2019). All samples were collected from Al-Diwanyia city. The results of PCR testing revealed that the primer pairs were specific and non-specific products did not appear for all samples. The amplification of Homo sapiens mitochondrial DNA with primer pairs of other (Ovis aries, Capra hircus, and Bos taurus) and amplification of each with primer pairs of another genus gave negative results, and this is primary evidence for primer pair specificity. The amplicon of 16S rRNA gene of Homo sapiens was 1200 bp, Ovis aries was 1060 bp, Capra hircus was 820 bp, and Bos taurus was 1300 bp. The sequencing revealed that no cross-reactivity of designed primer pairs and the PCR assay based on the designed primer pairs will be simple, fast, sensitive, specific, and cost-effective. There is sensitivity, specificity, and accuracy in the designed species-specific primer pairs and applicability of the designed primer pairs in forensics to investigate blood spots or evidence belonging for human, sheep, goat, and cow.

With the recent advances in high throughput next-generation sequencing technologies and bioinformatics approach, gut microbiome research, especially in livestock species, has expanded immensely, elucidating the greatest potential to investigate the unacknowledged understanding of rumen microbiota in host physiology at the molecular level. The association of a complex aggregated community of microbes to host metabolism is of great importance due to their crucial participation in metabolic, immunological, and physiological tasks. The knowledge of this sophisticated network of a symbiotic association of gut microbiota to host organisms may lead to novel insights for improving health, enhancing production, and reducing the risk of disease progression in livestock species necessary to meet the demands of the human race. The full picture of microorganisms present in a particular area can be achieved with the help of culture-independent omics-based approaches. The integration of metagenomics, metatranscriptomics, metaproteomics, and meta-metabolomics technologies with systems biology emphasizes the taxonomic composition, identification, functional characterization, gene abundance, metabolic profiling, and phylogenetic information of microbial population along with the underlying mechanism for pathological processes and their involvement as probiotic. The rumen secretions or partially digested feed particles, as well as fecal samples, are generally employed for gut microbiome investigation. The 16S rRNA gene sequencing amplicon-based technology is the most employed technique for microbiome profiling in livestock species to date. The use of software and biological databases in the field of gut microbiome research gives an accurate in-depth analysis of the microbial population greatly.

Research objectives : Identify genetic potential and community structure of soil and rhizosphere microbial community structure as affected by treated wastewater (TWW) irrigation. This objective was achieved through the examination soil and rhizosphere microbial communities of plants irrigated with fresh water (FW) and TWW. Genomic DNA extracted from soil and rhizosphere samples (Minz laboratory) was processed for DNA-based shotgun metagenome sequencing (Green laboratory). High-throughput bioinformatics was performed to compare both taxonomic and functional gene (and pathway) differences betwee