Saccharomyces cerevisiae S288C
Protocols: Haploid and diploid strains of Saccharomyces cerevisiae derived from S288C were used in the systematic deletion project. Haploid deletion mutant strains were constructed by substituting the candidate SUT/CUT locus with the kanMX4 cassette in BY4741 (MATa). Diploid deletion mutant strains were constructed in BY4743 (MATα) as a background which was generated from a cross between BY4741 (MATa) and BY4742 (MATα). Diploid deletion strains were constructed by substituting the candidate SUT/CUT locus with the kanMX4 cassette in BY4743 (MATa). Triplicates cultures (50 mL) of each ncRNA mutant strain and WT strain were grown in YPD to an OD600 of 0.5 (log phase) with shaking at 30˚C for RNA extraction. Total RNA was isolated using the RNeasy Mini Kit (QIAGEN, Germany) following the protocol for enzymatic digestion of cell wall followed by lysis of spheroplasts. To eliminate genomic DNA contamination, an additional DNAse treatment was performed with RNAse-free DNase set (QIAGEN, Germany) following the manufacturer's protocol. The RNA extracted was quantified using a NanoDrop LiTE Spectrophotometer (THERMO SCIENTIFIC, United States). Libraries were prepared from total RNA using the TruSeq Stranded mRNA Library Prep Kit (Illumina,Inc) according to the manufacturer's instructions.
ENA first public
2021-01-19
ENA last update
2020-12-21
External Id
SAMEA7745684
INSDC center alias
Bioinformatics Core Facility University of Mancheste
INSDC center name
Bioinformatics Core Facility University of Mancheste
INSDC first public
2021-01-19T17:07:00Z
INSDC last update
2020-12-21T22:06:58Z
INSDC status
public
Submitter Id
E-MTAB-9956:Sample 13
broker name
ArrayExpress
genetic modification
SUT532
genotype
haploid MATa ura30 leu20 his31 met150 SUT532::kanMX4
sample name
E-MTAB-9956:Sample 13