title: GSM3307586: Control-5; Mus musculus; RNA-Seq
library_construction_protocol: Testes were isolated, total RNA was isolated using RNeasy Mini Kit and mRNA was purified by Dynabeads mRNA Purification Kit. First strand cDNA was synthesized with SuperScript III Reverse Transcriptase. For second strand cDNA synthesis, the 100 μl reaction system included: 20 μl of first strand cDNA synthesis mix, 10 μl of 10x second strand buffer (500 mM Tris-HCl, pH 7.5; 50 mM MgCl2; 10 mM DTT), 3 μl of dNTP mix (10 mM), 1 μl of RNase H (2U/μl), 5 μl of DNA Pol I (10U/μl) and 61 μl water. The mixtures were incubated for 2 h at 16°C. The libraries were constructed with Nextera DNA Sample Preparation Kit (Illumina) per the manufacturer's protocol. Generally, double-strand cDNA was fragmented, subjected to adapter ligation and amplified.
attributes.tag: GEO Accession
attributes.value: GSM3307586