16s rRNA gene of mus musculus: gut microbiota
Bacterial DNA from fecal samples was isolated with QIAamp DNA Stool Mini Kits (Qiagen) and bacterial strains were investigated using 16S rRNA gene sequencing. 15 ng of DNA was used as template to amplify 16S rRNA gene using High Fidelity PCR system kit (Roche). The v1-v3 regions of 16S ribosomal RNA gene was amplified using 27f (AGAGTTTGATCCTGGCTCAG) and 534r (ATTACCGCGGCTGCTGG) primers (1 ?M). The primers were anchored with adaptor (adopter A: 5' CCATCTCATCCCTGCGTGTCTCCGACTCAG 3' and adopter B: 5' CCTATCCCCTGTGTGCCTTGGCAGTCTCAG 3') and Multiplex Identifiers (MIDs; 10 bp long). The multiplexed amplicons were purified using QIAquick Gel Extraction Kit (Qiagen).
SUB3156304
JC22