organism: Streptomyces albus J1074
description: Protocols: Streptomyces albus DEL_pseB4 strain described by Bilyk and Luzhetskyy, 2014. For total RNA isolation, S. albus cells were grown in NL19 medium (for indigenous BGC expression). Then, 2 ml of 2-day and 3-day cultures was spun down for 20 s at 14,000 × rpm, and the pellets were immediately frozen in liquid nitrogen and stored at ?80 °C. S. albus cells were grown in SG medium (for ara expression) or NL19 medium (for indigenous BGC expression). Total RNA extraction was performed using an RNeasy Kit (Qiagen, Hilden, Germany) as previously described (Huser et al. 2003). An RNase-Free DNase set (Qiagen) was used two times for on-column DNA digestion, and an additional DNase treatment was then performed with a DNase I kit (Roche Diagnostics, Mannheim, Germany) to ensure that all DNA was completely removed. To check the RNA samples for DNA contamination, PCR was performed using oligonucleotides designed to create two different products approximately 150 bp and 500 bp in size. Initially, RNA quality was checked with Trinean Xpose (Gentbrugge, Belgium) and Agilent RNA Nano 6000 kits on an Agilent 2100 Bioanalyzer (Agilent Technologies, Böblingen, Germany). A Ribo-Zero rRNA Removal Kit for bacteria was obtained from Illumina (San Diego, CA, USA) and used to remove ribosomal RNA molecules from isolated total RNA. rRNA removal was checked using an Agilent RNA Pico 6000 kit on an Agilent 2100 Bioanalyzer (Agilent Technologies, Böblingen, Germany). A TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA, USA) was used to prepare cDNA libraries (Koepff et al. 2017).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-05-01T04:02:51Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-02-19T15:35:34Z
attributes.tag: External Id
attributes.value: SAMEA5360516
attributes.tag: INSDC center name
attributes.value: Bielefeld University, Centre for Biotechnology (CiBiTec), Universitatsstraße 27, 33615, Bielefeld, Germany
attributes.tag: INSDC first public
attributes.value: 2019-05-01T04:02:51Z
attributes.tag: INSDC last update
attributes.value: 2019-02-19T15:35:34Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7715:KE-3day
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: genetic modification
attributes.value: K88E mutation in rpsL gene
attributes.tag: sample name
attributes.value: E-MTAB-7715:KE-3day
attributes.tag: scientific_name
attributes.value: Streptomyces albus J1074
attributes.tag: strain
attributes.value: DEL_pseB4