title: Pto+DMSO
organism: Pseudomonas syringae pv. tomato
description: Protocols: Pto DMSO, Pto +Ft and Pto +Na cells were harvested at OD660=1.5, washed with sarkosyl 0.1% and cell pellets were immediately frozen in liquid nitrogen and conserved at -80ºC until RNA isolation. Pseudomonas syringae pv. tomato (Pto) DC3000 was grown in LB at 28ºC to an OD660=0.5 in the absence and in the presence of phloretin and naringenin.Bacteria were grown on LB plates for 48 hours and resuspended in 3 ml of 10 mM MgCl2. Ten ml of LB with 0.01% DMSO, 0.5 mM phloretin or 0.5 mM naringenin (both in DMSO) were inoculated and adjusted to an OD660 of 0.05. The strains were grown at 28ºC to an OD660=0.5. rRNA removal with Ribo zero magnetic kit was performed by Macrogen. The cells were processed for RNA isolation using TRI Reagent®LS (Molecular Research Center, Cincinnati, OH, USA) according to the manufacturer instructions, except that the TRI Reagent was preheated at 70ºC and the lysis step was carried out at 65ºC. The RNA obtained was treated with RNase-free DNase I (Quiagen) followed by purification with RNeasy mini kit (Quiagen). RNA concentration was determined spectrophotometrically and its integrity was assessed by agarose gel electrophoresis. cDNA libraries were constructed by Macrogen
attributes.tag: ENA first public
attributes.value: 2018-12-30
attributes.tag: ENA last update
attributes.value: 2018-01-26
attributes.tag: External Id
attributes.value: SAMEA104549013
attributes.tag: INSDC center alias
attributes.value: Estacion Experimental del Zaidin (CSIC)
attributes.tag: INSDC center name
attributes.value: Estacion Experimental del Zaidin (CSIC)
attributes.tag: INSDC first public
attributes.value: 2018-12-30T17:01:47Z
attributes.tag: INSDC last update
attributes.value: 2018-01-26T09:51:21Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6426:Pto+DMSO
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: growth condition
attributes.value: liquid medium
attributes.tag: medium
attributes.value: LB
attributes.tag: optical density measurement
attributes.value: 0.5 OD at 660 nm
attributes.tag: replicate
attributes.value: 2
attributes.tag: sample name
attributes.value: E-MTAB-6426:Pto+DMSO
attributes.tag: strain
attributes.value: DC3000
attributes.tag: temperature
attributes.value: 28
attributes.tag: pathovar
attributes.value: tomato
attributes.tag: type-material
attributes.value: reference strain of Pseudomonas syringae group genomosp. 3