title: GSM5468485: 39-naivemdxT16VIVO06DMD-2018_R1; Mus musculus; miRNA-Seq
library_construction_protocol: RNA extraction from plasma sample was performed using miRNeasy Serum/Plasma Kit (Qiagen) following the manufacturer's instructions. Libraries for small RNA sequencing were prepared using the NEBNext small RNA library prep kit (New England Biolabs) according to the manufacturer's instructions. Briefly, 5 μl of RNA eluate was used as input for RNA adapter ligation (using 3' and 5' RNA adapters) followed by reverse transcription and PCR amplification with bar-coded primers. Size selection of individual libraries was performed on a Pippin Prep system (Sage Science) to recover the ~147-157 nt fractions containing mature miRNAs. Next, qPCR-based normalization was used for pooling of the individual small RNA libraries. Finally, the resulting small RNA pools were concentrated via ethanol precipitation and quantified using the Qubit 2.0 fluorometer (Thermo Fisher Scientific) prior to sequencing with read length of 75 bp on a NextSeq 500 sequencer (Illumina).