title: 16S rRNA sequencing of the musk microbiota from 5 unmated 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 the sequencing of the musk microbiota from the 5 unmated males. There is one BioSample and the BioSample has 5 biological replicates, each replicate is a sample from a unmated male, we sequenced the 5 replicates, respectively. Each experiment has one Run and there is one file in the Run.   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 ampliﬁed 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 ampliﬁed 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.