Bos taurus miRNA profile after DNA transfection - IncE
Cooled semen from seven bulls were centrifuged twice for 5 min at 700 g to remove commercial diluent. An aliquot of 100 x 106 cells from the sperm pellet was added in each treatment and used in subsequent analysis. The experimental groups were defined according to the method of exogenous DNA transfection carried out: incubation with (transfection) or without naked exogenous DNA (control) and Cationic dendrimer mediated transfection with exogenous DNA (dendriplex  Polyfect Transfection Reagent). Total RNA was isolated from sperm using a combined method of TRIzol Reagent and RNeasy mini spin column. Small RNA libraries were generated using NEB Next Small RNA Library Prep Set for Illumina and quantified by Agilent Technologies BioAnalyzer 2100. The libraries concentrations were determined using the Invitrogen Qubit dsDNA High Sensitivity Assay Kit and subsequently pooled at equimolar concentrations prior to Illumina MiSeq sequencing with 50-bp single-end reads configuration using MiSeq Reagent Kit v3 sequencing chemistry