Baseline_expression_from_transcriptional_profiling_of_zebrafish_developmental_stages. Baseline_expression_from_transcriptional_profiling_of_zebrafish_developmental_stages

Establish a baseline expression transcriptional signatures for zebrafish developmenal stages. Total RNA was extracted from pools of wild type zebrafish embryo at a series of different developmental stages in replicate. The 3’ end of fragmented RNA was pulled down using a 30T polyT oligo attached to magnetic beads, reverse transcribed, made into Illumina libraries and sequenced using Illumina HiSeq paired-end sequencing. Protocol: Total RNA was extracted from zebrafish embryo pools and DNase treated. Fragmented RNA was enriched for the 3’ ends by pulled down using an anchored 30 T polyT oligo attached to magnetic beads. An RNA oligo comprising part of the Illumina adapter 1 was ligated to the 5’ end of the captured RNA and the RNA was eluted from the beads. Reverse transcription was primed with an anchored polyT oligo with part of Illumina adapter 2 at the 5’ end followed by the 10 bases HBDVHBDVHB, then one of 96 eight base indexing tags, then CG and 14 T bases. An Illumina library with full adapter sequence was produced by 20 cycles of PCR. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing