2024-09-16
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114498151
External Id
Lorraine University
INSDC center name
2023-10-14T20:40:47Z
INSDC last update
public
INSDC status
ERS16479806
SRA accession
HPS rpa34DeltaK_rep3
Submitter Id
not provided
collection date
baker's yeast
common name
Total RNA from S.cerevisiae strain Yeast cells (wt, pxr1-?KK, tma23-?KK, rpa34?K, Nop56/58??K, cbf5?K) were grown exponentially at 30�C in YPD (1% yeast extract, 2% peptone, 2% glucose) medium or YPG (1% yeast extract, 2% peptone, 2% galactose) medium (WTGAl, rdn1?. pGal::rDNA. rpa135?) prior to rRNA extraction. Extraction of yeast total RNA: dry cell pellets were resuspended with 0.5?ml water-saturated phenol and 0.5?ml guanidine thiocyanate (GTC) mix (50?mM Tris-HCl, pH 8.0, 10?mM EDTA, pH 8.0, 4?M guanidine thiocyanate, 2% N-Lauroylsarcosine, 143?mM ?-Mercaptoethanol). Cells were broken by vigorous vortexing three times for 2?min at 4?�C in the presence of Zirconium beads. The resulting samples were mixed with 7.5?ml water-saturated phenol and 7.5?ml GTC mix and incubated for 5?min at 65?�C. After addition of 7.5?ml chloroform and 4?ml sodium acetate buffer (10?mM Tris-HCl, pH 8.0, 1?mM EDTA, pH 8.0, 100?mM sodium acetate), samples were mixed vigorously and centrifuged at 3220?�?g for 5?min at 4?�C. Aqueous phases were recovered and RNAs were re-extracted three additional times with water-saturated phenol:chloroform (1:1). RNAs were then concentrated by ethanol precipitation and ultimately resuspended in ultrapure H2O.
description
France
geographic location (country and/or sea)
Saccharomyces cerevisiae
organism
Saccharomyces cerevisiae
scientific_name
rpa34DeltaK_rep3
title