16S rRNA based microbiome analysis of Homo sapiens: healthy lactating North Indian mothers
16S rRNA gene (V1-V5) was amplified in a 50-l reaction volume using Q5 Taq polymerase (NEB, USA), 0.1 ng of template DNA and barcoded forward (27F) and reverse (926R) primers. The amplified PCR products (950-bp) were purified using gel elution Kit (Qiagen, Germany). Equimolar concentration of purified DNA libraries from all the samples were sequenced using 454 GS FLX+ pyrosequencer (Roche, Switzerland).