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10.1038/s41598-018-23296-4 http://scihub22266oqcxt.onion/10.1038/s41598-018-23296-4 C5865204!5865204 !29572539     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=29572539 &cmd=llinks): Failed to open stream: HTTP request failed! 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Methodology challenges in studying human gut microbiota - effects of collection, storage, DNA extraction and next generation sequencing technologies |pdf=|usr=}}{{29572539 }} gab.com Text Methodology challenges in studying human gut microbiota - effects of collection, storage, DNA extraction and next generation sequencing technologies JO: Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=29572539 #FOAvip The information on microbiota composition in the human gastrointestinal tract predominantly originates from the analyses of human faeces by application of next generation sequencing (NGS). However, the detected composition of the faecal bacterial community can be affected by various factors including experimental design and procedures. This study evaluated the performance of different protocols for collection and storage of faecal samples (native and OMNIgene.GUT system) and bacterial DNA extraction (MP Biomedicals, QIAGEN and MO BIO kits), using two NGS platforms for 16S rRNA gene sequencing (Ilumina MiSeq and Ion Torrent PGM). OMNIgene.GUT proved as a reliable and convenient system for collection and storage of faecal samples although favouring Sutterella genus. MP provided superior DNA yield and quality, MO BIO depleted Gram positive organisms while using QIAGEN with OMNIgene.GUT resulted in greatest variability compared to other two kits. MiSeq and IT platforms in their supplier recommended setups provided comparable reproducibility of donor faecal microbiota. The differences included higher diversity observed with MiSeq and increased capacity of MiSeq to detect Akkermansia muciniphila, [Odoribacteraceae], Erysipelotrichaceae and Ruminococcaceae (primarily Faecalibacterium prausnitzii). The results of our study could assist the investigators using NGS technologies to make informed decisions on appropriate tools for their experimental pipelines. Twit Text FOAVip Methodology challenges in studying human gut microbiota - effects of collection, storage, DNA extraction and next generation sequencing technologies ????Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=29572539 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=29572539 #FOAvip English Wikipedia <ref>{{Cite pmid|29572539 }}</ref> Methodology challenges in studying human gut microbiota - effects of collection, storage, DNA extraction and next generation sequencing technologies #MMPMID29572539 Panek M ; ?ip?i? Paljetak H ; Bare?i? A ; Peri? M ; Matija?i? M ; Lojki? I ; Vrane?i? Bender D ; Krznari? ? ; Verbanac D Sci Rep 2018[Mar]; 8 (1 ): 5143 PMID29572539 show ga The information on microbiota composition in the human gastrointestinal tract predominantly originates from the analyses of human faeces by application of next generation sequencing (NGS). However, the detected composition of the faecal bacterial community can be affected by various factors including experimental design and procedures. This study evaluated the performance of different protocols for collection and storage of faecal samples (native and OMNIgene.GUT system) and bacterial DNA extraction (MP Biomedicals, QIAGEN and MO BIO kits), using two NGS platforms for 16S rRNA gene sequencing (Ilumina MiSeq and Ion Torrent PGM). OMNIgene.GUT proved as a reliable and convenient system for collection and storage of faecal samples although favouring Sutterella genus. MP provided superior DNA yield and quality, MO BIO depleted Gram positive organisms while using QIAGEN with OMNIgene.GUT resulted in greatest variability compared to other two kits. MiSeq and IT platforms in their supplier recommended setups provided comparable reproducibility of donor faecal microbiota. The differences included higher diversity observed with MiSeq and increased capacity of MiSeq to detect Akkermansia muciniphila, [Odoribacteraceae], Erysipelotrichaceae and Ruminococcaceae (primarily Faecalibacterium prausnitzii). The results of our study could assist the investigators using NGS technologies to make informed decisions on appropriate tools for their experimental pipelines. |*DNA, Bacterial/genetics/isolation & purification [MESH] |*Gastrointestinal Microbiome [MESH] |*Gram-Positive Bacteria/chemistry/classification/genetics/isolation & purification [MESH] |*High-Throughput Nucleotide Sequencing [MESH] |Adult [MESH] |Feces/*microbiology [MESH] |Female [MESH] |Humans [MESH] |Male [MESH] |Reproducibility of Results [MESH]Methodology challenges in studying human gut microbiota - effects of c(epmc).pdf DeepDyve Pubget Overpricing