title: 16S rRNA sequencing of the musk microbiota from one of the mated Chinese forest musk deer       males
design: ? We sequenced the musk microbiota of 5 unmated and 5 mated Chinese forest         musk deer males by using the 16S rRNA sequencing method on the Illumina Miseq platform in         early December 2014, the Chinese forest musk deer males were provided by Chongqing Institute         of Medicinal Plant Cultivation (Chongqing, China). This is one of the sequenced musk         microbiota from the mated male. ? There is two BioSampls and each BioSample has 5 biological         replicates, each replicate has one experiment, each experiment has one Run, respectively,         and each Run has two files (FASTA and QUAL). ? Sequencing methods: We used Illumina Miseq         platform to sequence the 16S rRNA of the musk microbiota and performed multiplex         pyrosequencing of the V3-V4 hyper-variable regions of 16S rRNA gene. ? Kits used: Total         bacteria DNA was extracted from musk samples by using MO BIO UltraClean fecal DNA kit         (Carlsbad, CA, USA). Bacterial 16S rRNA genes in musk were PCR ampli?ed by using two broadly         conserved, degenerate primer set targeting the V3–V4 variable regions of the 16S gene. ?         Instrument model: PCR reaction was conducted using a Mastercycler Gradient Thermal Cycler         (Eppendorf) with the following condition: 94 °C for 3 min (initial dissociation), followed         by 30 cycles of [94 °C for 30 s (denaturing), 60 °C for 45 s (annealing), and 72 °C for 1         min (extending)], followed by a final extension of 8 minutes at 72°C. Non template controls         were run for each barcoded primer pair. Each sample was ampli?ed and then pooled together         for analysis. The PCR product was analyzed by gel electrophoresis and purified by using a         PCR Purification kit (Sangon, Shanghai, China). Pyrosequencing was performed on an Illumina         MiSeq 2 × 250 platform.