Microbial Composition and Co-Occurrence Patterns in the Gut Microbial Community of Normal and Obese Mice in Response to Astaxanthin

The microbial genomic DNA from the colon
contents of NO, NA, MO and MA groups were extracted with a QIAamp DNA stool Mini
kit (Qiagen, cat.
No 51604). The 16S rRNA sequencing and analysis was used
for obtaining the gut microbiota profiling. The V3-V4 regions of the 16S rRNA
gene were selected for amplification. The total DNA were amplified with specific
primers (forward primer, 341/357F:
NNNNCCTACGGGNGGCWGCAG; reverse primer, 805/785R: GACTACHVGGGTATCTAATCC) tagged
with barcodes. The PCR reactions were carried out with Phusion
high-fidelity PCR Master Mix (New England Biolabs, Ipswich, MA, USA). The
concentration of the library pool was quantified by a BioAnalyzer DNA chip kit
(Agilent) and the library was sequenced by an Illumina sequencer.