ena-STUDY-UNIVERSITY OF ABERDEEN-04-10-2018-11:47:49:434-50
Cyba-nmf333 mice have altered gut microbiota and their increased risk of colitis is abolished by Streptomycin treatment.
This study includes two related experiments: (1) analysis of adherent and luminal colon microbiota from wild type (WT) and Cybanmf333 mice (n=8-10); (2) analysis of faecal pellet microbiota from wild type and Cybanmf333 before (WTdm1 and Cybanmf333dm1) and after (WTd0 and Cybanmf333d0)Streptomycin treatment (n=9-8). Mouse groups included males and females, aged 10-16 weeks and house in separate cages. Streptomycin (1g/L) in drinking water was given for two days. DNA was isolated using QIAamp DNA Stool Mini Kits (Qiagen). DNA concentration and purity was monitored on 1% agarose gels. The V4 16S rRNA gene region was PCR amplified using primers V4-515F and V4-806R with barcodes by Novogene (Hong Kong). All PCR reactions were carried out with Phusion® High Fidelity PCR Master Mix (New England Biolabs). Sequencing libraries were generated using TruSeq® DNA PCR-Free sample preparation kit (Illumina, USA) following the manufacturer's recommendations, and index codes were added. The library was then sequenced on an IlluminaHiSeq2500 platform and 250 bp paired-end reads were generated (Novogene, Hong Kong).
This study includes two related experiments: (1) analysis of adherent and luminal colon microbiota from wild type (WT) and Cybanmf333 mice (n=8-10); (2) analysis of faecal pellet microbiota from wild type and Cybanmf333 before (WTdm1 and Cybanmf333dm1) and after (WTd0 and Cybanmf333d0)Streptomycin treatment (n=9-8). Mouse groups included males and females, aged 10-16 weeks and house in separate cages. Streptomycin (1g/L) in drinking water was given for two days. DNA was isolated using QIAamp DNA Stool Mini Kits (Qiagen). DNA concentration and purity was monitored on 1% agarose gels. The V4 16S rRNA gene region was PCR amplified using primers V4-515F and V4-806R with barcodes by Novogene (Hong Kong). All PCR reactions were carried out with Phusion® High Fidelity PCR Master Mix (New England Biolabs). Sequencing libraries were generated using TruSeq® DNA PCR-Free sample preparation kit (Illumina, USA) following the manufacturer's recommendations, and index codes were added. The library was then sequenced on an IlluminaHiSeq2500 platform and 250 bp paired-end reads were generated (Novogene, Hong Kong).