delPse-2day
Streptomyces albus J1074
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).
ENA-FIRST-PUBLIC
2019-05-01T04:02:51Z
ENA-LAST-UPDATE
2019-02-19T15:35:33Z
External Id
SAMEA5360509
INSDC center name
Bielefeld University, Centre for Biotechnology (CiBiTec), Universitatsstraße 27, 33615, Bielefeld, Germany
INSDC first public
2019-05-01T04:02:51Z
INSDC last update
2019-02-19T15:35:33Z
INSDC status
public
Submitter Id
E-MTAB-7715:delPse-2day
broker name
ArrayExpress
genetic modification
none add. to Del_pseB4
sample name
E-MTAB-7715:delPse-2day
scientific_name
Streptomyces albus J1074
strain
DEL_pseB4