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Fecal sample collection and 16s rRNA Illumina sequencing

KA Kazi Farhana Afroz
NR Noah Reyes
KY Kobe Young
KP Kajal Parikh
VM Varsha Misra
KA Karina Alviña
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Colonic fecal samples were collected from the dams of both diet groups before weaning. The fecal samples from the offspring groups were collected before weaning at 2–3 weeks of age (i.e. while they were still housed with their mothers). To reduce stress, mice were placed inside autoclaved plastic boxes where they freely defecated. Feces were then collected and placed in autoclaved cryotubes, flash frozen in liquid nitrogen and immediately stored at − 80 °C. DNA extraction from the samples, Polymerase Chain Reaction (PCR) amplification of the variable 4 (V4) region of the 16s rRNA gene and sequencing using the Illumina MiSeq platform were performed by the Molecular Research LP laboratories (MR DNA, Shallowater, TX, USA).

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