title: GSM3553950: HighGFP_1; Danio rerio; RNA-Seq
library_construction_protocol: 10 Optic vesicles each were dissected manually from about 18 ss wild-type (control) embryos and Tg(vsx2::GFP) embryos. The dissected optic vesicles were kept in 100 μl PBS at room temperature. The cells were dissociated manually by pipetting several times and the volume was adjusted to 300 μl. This procedure was performed within half an hour and samples were used for FACS. For the RNA-seq experiment, the SMARTer Ultra Low Input RNA for Illumina Sequencing Kit - HV from Takara Bio USA (formerly known as Clontech Laboratories) was used to reverse transcribe the RNA and amplify full-length cDNA according to the user manual. After amplification of cDNA, the cDNA was sheared to a fragment length of 200 bp in the ultrasonicator Covaris LE220, followed by a library prep using NEBNext® Ultra DNA Library Prep Kit for Illumina (NEB). The resulting libraries were pooled and sequenced on an Illumina® HiSeq 2500, resulting in ca. 25 – 60 million single-end reads.
attributes.tag: GEO Accession
attributes.value: GSM3553950