GSM1155235
GSM1155235: (-)4SUTP_512cell; Danio rerio; RNA-Seq
Total RNA from staged embryos was extracted with Trizol, biotinylated with EZ-Link HPDP-Biotin and subsequently purified with streptavidin-coated magnetic beads to isolate newly transcribed RNA. Labeled and unlabeled samples were treated in the same way. Libraries were prepared for the HiSeq 2000 machine (Illumina). Briefly, RNA was amplified with the WT-Ovation™ One-Direct RNA Amplification System (NuGEN). The reaction was stopped after the SPIA amplification and double-stranded cDNA with random Hexamers created. Double-stranded cDNA was sheared to ~200-300bp fragment size, end-repaired with the NEBNext End Repair Module (NEB) and the resulting fragments were adenylated at the 3'-end with the NEBNext dA-Tailing Module (NEB).TruSeq adapters were ligated and the libraries were PCR amplified for 15 cycles. The concentration of molecules with adapters ligated in the final library was determined by qPCR with the KAPA Library Quant Kit (Kapa Biosystems).
GEO
GSM1155235
GEO Accession
GSM1155235