GSM5076754: F1N_D_RNA_6; Mus musculus; ncRNA-Seq
RNA from purified sperm (centrifugation) was extracted using TRIzol reagent. The quality of the RNA preparations was verified by spectrometry on an Agilent Bioanalyzer 2100 apparatus. Microinjection into fertilized eggs was performed according to established transgenesis methods. RNA solutions were adjusted to a concentration of 1 µg/ml, and 1-2 pl were microinjected into the male pronuclei of fertilized eggs isolated after normal ovulation and mating to a Balb/c strain. Synthetic miRNA was prepared in a filtered microinjection buffer (10 mM Tris, pH 7.4; 0.1 mM EDTA) at a concentration of 3,000,000-5,000,000 molecules/pl. After digestion of DNase, 10-100 ng of RNA were prepared and sent to Eurofins for high-throughput sequencing on Illumina Hiseq 2500 or Illumina MiSeq. Libraries of all samples corresponding to DNA-associated RNA and free-RNA molecules were generated