GSM2801058: Neonatal-3; Mus musculus; ncRNA-Seq
Ovaries were removed, flash frozen on dry ice, and RNA was harvested using TRIzol reagent (Life Technologies, CA, USA), total RNA (5μg) was incubated for 15 min at 37°C with 3U/μg of RNase R (Epicentre Biotechnologies)  was used with 1 ug of total RNA for the construction of sequencing libraries. Then the rRNA depleted RNA samples or circRNAs samples were fragmented and used to synthesize first- and second-strand complementary DNA (cDNA) with random hexamer primers, dNTPs and DNA Polymerase I. The cDNA fragments were cleaned and concentrated using AMPure XP beads, then the ends were repaired and modified with T4 DNA polymerase Klenow DNA polymerase to add A and adapter at the 3’ end of the DNA fragments. The ligated cDNA products were purified with AMPure XP beads and treated with uracil DNA glycosylase to remove the second-strand cDNA. Purified first-strand cDNA was subjected to PCR amplification, and the libraries were quality controlled with an Agilent 2100 Bioanalyzer and sequenced by HiSeq 2500 (Illumina, San Diego, CA) on a 150 bp paired-end (PE) run.
GEO
GSM2801058