PARE-Seq of mitochondrial RNA from Ang+/+ fusing osteoclasts
Mitochodrial RNAs was extracted from mitochondria according to the protocol provided in the RNeasy Mini Kit (QIAGEN, 74014). The cleaved mitochondrial polyA-RNA samples were ligated to a single-stranded 5 RNA adaptor using RNA ligase, followed by reverse transcription and amplification. The PCR products were digested with MmeI, a type IIS restriction enzyme, to generate equal-sized fragments, which were recovered by PAGE. After ligation with a 3 adapter containing degenerate nucleotides in the overhang region, the digested RNAs were amplified and gel-purified for deep sequencing on the Illumina HiSeq SE50 by CD Genomics (Shirley, USA).
SUB16124189
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