GSM9320834: Female, Fecal sequence, HfiD feed 5w, HFD feed 0w; Mus musculus; OTHER
Fecal pellets were collected and stored at -80°C until DNA extraction. DNA extraction was performed using the Magnetic Soil and Stool DNA Kit (TianGen, Cat# DP712) and verified on a 2% agarose gel for band at around 310 bp. Primers 515F (GTGCCAGCMGCCGCGGTAA) and 806R (GGACTACHVGGGTWTCTAAT) with barcode were used for bacterial hypervariable V4 region amplification. Sequencing libraries were built with the NEB Next® Ultra™ II FS DNA PCR- free Library Prep Kit (New England Biolabs, Cat# E7430L) and validated with Qubit and real-time qPCR before sequencing with the NovaSeq 6000 system (Illumina).