16S rRNA sequencing of mouse cecal microbiota
The DNA was isolated from the given samples and the DNA concentration was estimated using Qubit Fluorimeter(V.3.0). The V3-V4 region of 16S rRNA was amplified using specific V3 Forward primer CCTACGGGNBGCASCAGand V4 Reverse primer GACTACNVGGGTATCTAATCC. The amplified productwas checked on 2% agarose gel and gel purification was done to remove non-specific amplifications.5ng ofamplified product was used for library preparation using NEBNext Ultra DNA library preparation kit. Thelibrary quantification and quality estimation were done in Agilent 2200 TapeStation. The prepared librarywas sequenced in Illumina HiSeq 2500 platform.