2026-10-12T06:45:28Z https://www.scidb.cn/oai

10.57760/sciencedb.013vu 2026-09-28T15:57:20Z

2026-09-28 SPFvsGF_5R16S rRNA doi:10.57760/sciencedb.013vu en Specified Pathogen Free (SPF, n=16) and germ free (GF, n=16) mice (C57BL/6) were purchased from Kunshan Mingqian Microbiology Research Institute（Kunshan, China）。 To prevent contamination of GF mice during transportation and maintain consistent sampling environments for both groups of samples, ovarian sampling was performed in a sterile manner in the laboratory's ultra clean workbench of the purchasing company. After ovarian collection, it was sealed and transported in dry ice to Hangzhou Lianchuan Biotechnology Co., Ltd. (Hangzhou, China) for 5R 16S rRNA gene sequencing. The DNA of ovarian tissue was extracted using a bacterial genome extraction kit (TIANGEN, DP302-02). Design primers targeting the variable regions of 16S rDNA V2, V3, V5, V6, and V8 (5R primers; Supplementary Table 1) (5R primers: V2 forward TGGGAACGGGTGTAAA, V2 reverse CCGTGTCTCAGTCCCARTG; V3 forward - ACTCCTACGGGAGGCAGC, V3 reverse - GTATTACCGCGGCTGCTG; V5 forward - GTGTAGGGTGRAATGCG, V5 reverse - CCCGTCAATTCMTTGAGTT; V6 forward - GGAGCATGTGGWTTAATTCGA, V6 reverse - CGTTGCGGGACTTAACCC; V8 Forward - GGAGGAAGGTGGGGATGAC, V8 Reverse - AAGGCCCGGGAACGTATT; ）Perform PCR amplification, purify and quantify the PCR products using AMPure XP beads (Beckman Coulter Genomics, Danvers, MA, USA), and perform high-throughput sequencing (NovaSeq 6000 SP Reagent Kit; Illumina NovaSeq 6000). The Short Multi Regions Framework (SMURF) was used to integrate sequencing data from five variable regions, and the 16S rRNA fragments were reconstructed using the maximum expected algorithm. Species taxonomy annotation was performed based on the Greengenes2 (2024) database.  SPF mice; GF mice; 5R 16S rRNA gene sequencing; ovary; Ovarian microecology Liu Zhenxing EMBARGO https://creativecommons.org/licenses/by/4.0/ dataset Science Data Bank