title: eIF4E-Total(1)
organism: Saccharomyces cerevisiae
description: Protocols: Grown at 30C overnight to an OD600 0.6, pelleted by centrifugation and snap frozen into liquid nitrogen Affinity chromatography is used to pulldown the protein of interest with attached mRNAs. The protein of interest has a protein A tag encoded on the genome of the yeast strain and IgG (the same antibody is used across all samples) magnetic beads are used to pull out this protein from whole cell extracts. The RNA is processed from these samples and compared to total RNA samples from the same strains. samples ground under liquid nitrogen in lysis buffer using a 6870 Freezer Mill (Spex). Lysate thawed on ice, clarified by centrifugation and nucleic acids extract using Trizol Reagent. rRNA was depleted from samples using Ribominus Eukaryote Kit, ethanol precipitated and resuspended in DEPC water. Sequencing libraries were then generated using the whole Transcriptome Library Preparation protocol provided with the SOLiD Total RNA-Seq Kit. . cDNA was then amplified and barcoded with SOLiD, RNA barcoding Kit and samples purified using PureLink, PCR Micro Kit
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2014-08-01T17:01:08Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T17:21:14Z
attributes.tag: External Id
attributes.value: SAMEA2465344
attributes.tag: INSDC center name
attributes.value: The University of Manchester
attributes.tag: INSDC first public
attributes.value: 2014-08-01T17:01:08Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T17:21:14Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2464:eIF4E-Total(1)
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: baker's yeast
attributes.tag: genotype
attributes.value: eIF4E-tap-tag
attributes.tag: sample name
attributes.value: E-MTAB-2464:eIF4E-Total(1)
attributes.tag: scientific_name
attributes.value: Saccharomyces cerevisiae
attributes.tag: sex
attributes.value: Mat a
attributes.tag: specimen with known storage state
attributes.value: fresh specimen
attributes.tag: strain
attributes.value: BY4741