title: I56T_SS_SORB
organism: mixed sample
description: Protocols: Derivatives of Pichia pastoris strain GS115 were grown to steady state in chemostat cultures (1L volume in 2L fermenter; 30oC; pH5; 750 rpm stirring; air flow 1L/min) in a defined carbon-limited medium based on sorbitol (20 g/L). Dilution rates at steady state were 0.004-0.015 L/h, and all other non-carbon nutritional requirements were in excess at constant residual concentrations. Heterologous protein synthesis was induced in the strains by changing the carbon source composition of the medium to sorbitol (11.3 g/L) plus methanol (9 g/L). Samples were taken immediately prior to changing to the methanol media, 1 hr and 3 hr after methanol induction and finally after reaching a second steady state in the new media. Cells were lysed mechanically at 4oC in the presence of trizol. After partitioning with chloroform, RNA was precipitated from the aqueous phase using isopropanol. RNA was washed with 70% ethanol, and redissolved in water. 10 microg aliquots of the final RNA solution were ethanol precipitated.
attributes.tag: CellLine
attributes.value: GS 115
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2013-09-12
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2013-11-04
attributes.tag: External Id
attributes.value: SAMEA2007349
attributes.tag: Genotype
attributes.value: pPIC9-HuLy_I56T
attributes.tag: INSDC center alias
attributes.value: Cambridge Systems Biology Centre
attributes.tag: INSDC center name
attributes.value: Cambridge Systems Biology Centre
attributes.tag: INSDC first public
attributes.value: 2013-09-12T17:00:57Z
attributes.tag: INSDC last update
attributes.value: 2013-11-04T20:06:01Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-1126:I56T_SS_SORB
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: sample name
attributes.value: E-MTAB-1126:I56T_SS_SORB
attributes.tag: component_organism
attributes.value: Komagataella pastoris
attributes.tag: component_organism
attributes.value: Komagataella pastoris