title: 16S rRNA sequencing of the musk microbiota from one of the 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 one of the sequenced musk         microbiota from the 5 unmated males. ? There is two BioSamples and each has five         experiments, 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.