title: WT
organism: Schizosaccharomyces pombe
description: Protocols: We used a wild-type Schizosaccharomyces pombe 972 h- strain for all experiments. The strain was exponentially grown at 32 degree C in rich medium supplemented with glucose, adenine, histidine, lecucine and uracil. Cells were harvested and resuspended with pre-cooled lysis buffer (10mM EDTA, 10 mM Tris-HCl pH 8.0 and 50 mM NaOAc), which were further treated with mechanical pulverization. The cell material was stored at -80 degree C. Total RNA was extracted with the TRIZOL reagent according to the manufacturer's instructions (Invitrogen, Carlsbad, CA) and isolated RNA was quantified by bioanalyzer 2100. The CAGE library was prepared as the method described (Valen et al., Genome Res 2009) with some modifications to work with Illumina GA II sequencer. Briefly, cDNA synthesis was conducted by using 50ug total RNA and Superscript II revese transcriptase (Life Technologies). The cap selected cDNA/RNA hybrids were treated with RNase ONE Ribonuclease (promega) and purified cDNA was ligated with 5 prime end linker containing Mme I site. After 2nd strand synthesis, dsDNA fragments were digested with Mme I followed by the ligation with 3 prime end linker. The resulting CAGE tags were amplified by PCR, which was subject to deep sequencing.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2015-03-13T17:01:50Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T21:49:28Z
attributes.tag: External Id
attributes.value: SAMEA3177872
attributes.tag: INSDC center name
attributes.value: Shanghai Jiao Tong University
attributes.tag: INSDC first public
attributes.value: 2015-03-13T17:01:50Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T21:49:28Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-3188:WT
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: fission yeast
attributes.tag: growth condition
attributes.value: Normal
attributes.tag: sample name
attributes.value: E-MTAB-3188:WT
attributes.tag: scientific_name
attributes.value: Schizosaccharomyces pombe