Alteration of commensal microbiota by GP2- deficiency in the early age.

Pancreatic GP2 deficiency condition in early life of mice (2-3.5 weeks old) by administration of tamoxifen to pancreas-specific GP2 Floxed mice was made. Feces was collected at week 14 and examined by 16s rRNA gene sequencing. Bacterial DNA was extracted from the feces using an E.Z.N.A Stool DNA kit (Omega Bio-Tek, USA), and purified using magLEAD 12gc (Precision System Science, Japan). PCR was performed using KAPA HiFi HotStart ReadyMix (Nippon Genetics, Japan) and the primer set (Forward: 5-CCTNCGGGNGGCNGCAG-3, and Reverse: 5-GGATTAGATACCCN-NGTAGTC-3) to the V3-4 region of 16S rRNA gene. The amplicons were purified using AMPure XP (Beckman Coulter, USA). The DNA from each sample was added to different index sequence using Nextera XT index kit (Illumina, USA).