Transcriptome profiling of zebrafish biliary mutants
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ABSTRACT: Total RNA was extracted from morpholigcally abnormal and wildtype sibling zebrafish biliary mutants initially identified in a chemical mutagenesis screen. Mutants showed abnormal processing of the fluorescent lipid PED-6. The 3' end of fragmented RNA was pulled down using polyT oligos attached to magnetic beads, reverse transcribed, made into Illumina libraries and sequenced using IlluminaHiSeq paired-end sequencing. Protocol: Total RNA obtained was DNase treated. Chemically fragmented RNA was enriched for the 3' ends by pull down using an anchored polyToligo attached to magnetic beads. An RNA oligo comprising part of the Illumina adapter 2 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 polyToligo with part of Illumina adapter 1 at the 5' end followed by 4 random bases, then an A, C or G base, the one of twelve 5 base indexing tags and 14T bases. An Illumina library with full adapter sequence was produced by 15 cycles of PCR. 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 2000
ORGANISM(S): Danio rerio
SUBMITTER:
PROVIDER: E-ERAD-228 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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