Baseline expression from transcriptional profiling of zebrafish developmental stages

ABSTRACT: 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

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Danio rerio

SUBMITTER:  

PROVIDER: E-ERAD-274 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress