Polysome100uM_Farnesol3
Protocols: Grown in YPD media at 37oC to exponential phase (OD600 0.5-0.7), pelleted by centrifugation and snap frozen into liquid nitrogen. Some samples were treated with 100uM farnesol for 15minutes before RNA extraction. Samples lysed in Lola lysis buffer using a 6870 Freezer Mill (Spex). Lysates cleared by centrifuging twice at 15000g. For polysome associated samples 10A260 units were loaded onto a polysome gradient, fractionated then polysome fractions pooled. RNA was extracted from polysome associated and Total samples using Trizol Reagent followed by isopropanol precipitation. rRNA was depleted from samples using the Ribo-Zeroâ˘ Magnetic Gold Kit (Yeast) for RNA-Seq (Epicentre, Madison). Depleted samples were ethanol precipitated and re-suspended in 10 ď­l DEPC water. Sequencing libraries were generated using the TruSeq Stranded mRNA sample preparation kit from Illumina.
ENA first public
2016-07-31
ENA last update
2015-08-27
External Id
SAMEA3516272
INSDC center alias
UOM
INSDC center name
University of Manchester
INSDC first public
2016-07-31T17:03:02Z
INSDC last update
2015-08-27T16:46:28Z
INSDC status
public
Submitter Id
E-MTAB-3843_1:Polysome100uM_Farnesol3
broker name
ArrayExpress
fraction
polysome-associated RNA
genotype
ura3::imm434/ura3::imm434
sample name
E-MTAB-3843_1:Polysome100uM_Farnesol3