title: 130814puf3_RIP_1
organism: Saccharomyces cerevisiae
description: Protocols: Grow single colony overnight in 5ml YPD at 30C unless the growth media is otherwise specified in sample annotations. Dilute in 50-100ml desired medium with starting OD600=0.1. Grow to OD600=1.0. Harvest cells for total RNA extraction. Flash frozen pellets of exponentially growing TAP tagged cells (OD600 0.5-0.8) were mechanically lysed with glass beads using a FastPrep24 agitator. An aliquot of each cell lysate was used as an input control. Cell lysates were subjected to immunoprecipitation with Dyna M280 sheep anti-rabbit IgG beads (Invitrogen) for 2h at 4??C. RNA-protein complex was released by TEV protease cleavage of the TAP-tag. The immunoprecipitated RNA was purified by phenol:chloroform:isoamyl alcohol extraction. In summary 10 ug of total RNA was fragmented, purified, retrotranscribed, end-repaired, ligated, amplified and finally size selected. Full protocol is describe in An efficient method for genome-wide polyadenylation site mapping and RNA quantification Wilkening et al., 2013
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2014-11-13T17:02:20Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T16:54:04Z
attributes.tag: External Id
attributes.value: SAMEA2259376
attributes.tag: INSDC center name
attributes.value: European Molecular Biology Laboratory
attributes.tag: INSDC first public
attributes.value: 2014-11-13T17:02:20Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T16:54:04Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2103:130814puf3_RIP_1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: baker's yeast
attributes.tag: genotype
attributes.value: puf3-TAP
attributes.tag: sample name
attributes.value: E-MTAB-2103:130814puf3_RIP_1
attributes.tag: scientific_name
attributes.value: Saccharomyces cerevisiae
attributes.tag: sex
attributes.value: mating type alpha
attributes.tag: strain
attributes.value: BY4742 MAT alpha, his3-delta-1, leu2-delta-0, lys2-delta-0, ura3-delta-0, pep4-delta-::KANR