title: GSM1264048: 64hr - total; Ascaris suum; RNA-Seq
library_construction_protocol: RNA from early embryos were extracted directly with Trizol (Invitrogen). Frozen testis, ovary, oocyte, larvae, intestine, muscle, male carcass and female carcass samples were first ground with a mortar and pestle in liquid nitrogen and then extracted with Trizol. For each sample, 200 µg of total RNA was used for poly(A) selection and 200 ng of poly(A)+ RNA was used to make the cDNA libraries. RNA was treated for 75-150 sec with RNA fragmentation buffer (Ambion) at 70°C to generate RNA fragments 150-400 nt in length. First stand cDNA was prepared by random priming (3-6 µg random hexamers/1 µg of A+ RNA) and second stranded cDNA synthesis was carried out using a SuperScript® Double-Stranded cDNA Synthesis Kit (Invitrogen).  Solexa paired-end adaptors were added to the blunt cDNA as described by Illumina, cDNA fragments of 300-400 bp in length were gel purified, the fragments amplified using 17 PCR cycles, and sequencing reads were generated as described on the Illumina GAII, HiSeq2000, or MiSeq. For total and ribozero RNA (Ribo-Zero™ rRNA Removal Kits, Human/Mouse/Rat, Epicentre MRZH116), TotalScript RNA-seq kit (Epicentre, TSRNA1296) was used to generate the libraries.
attributes.tag: GEO Accession
attributes.value: GSM1264048