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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptome profiling of mutant embryos form the Mouse Genetics Project DMDD

ABSTRACT: Total RNA was extracted from embryonic lethal homozygous gene knockout and sibling embryos from the Mouse Genetics Project (http://www.dmdd.org.uk/). 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 Illumina HiSeq paired-end sequencing. Protocol: Total RNA was extracted from mouse embryos and DNase treated. Fragmented RNA was enriched for the 3 ends by pulled down using an anchored polyT oligo 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, then one of twelve 5 base indexing tags and 14 T bases. An Illumina library with full adapter sequence was produced by 15 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 2000

ORGANISM(S): Mus musculus

SUBMITTER:

PROVIDER: E-ERAD-237 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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