rrp41.1_rna
Schizosaccharomyces pombe
Protocols: S. pombe cells were grown in EMM medium supplemented with 20 ug/ml Thiamine at 30C for 10 hours (final OD600: 0.5-0.6). Total RNA was isolated from S. pombe cells using a hot phenol method followed by phenol-chloroform extractions, and precipitation as described (Lyne et al., 2003, BMC Genomics 4, 27.). 10ug total RNA was used as starting material for strand-specific RNA-seq library preparation using an unreleased early version provided by Illumina of the Illumina TruSeq Small RNA Sample Prep Kit. Briefly: poly(A)+ RNA was enriched by two rounds of poly(dT) Sera-Mag magnetic beads purification. Enriched poly(A)+ RNA was then fragmented to an average size of ~200nt. Fragmented RNA was 3' de-phosphorylated with Antartic phosphatase and 5' phosphorylated with polynucleotide kinase this treatment prepares RNA fragments for subsequent ligation of Illumina RNA adaptors to their 5' and 3' ends using a 3' RNA ligase and a T4 RNA ligase, respectively. First strand cDNA was produced using a primer specific for the Illumina 3' adaptor. The library was finally amplified with 15 cycles of PCR using primers specific for the Illumina adaptors and purified using SPRI-beads (Agencourt, Beckman Coulter). Libraries size distributions and concentrations were determined on an Agilent Bioanalyzer
ENA-FIRST-PUBLIC
2014-09-20T17:03:04Z
ENA-LAST-UPDATE
2018-03-08T17:08:16Z
External Id
SAMEA2335645
INSDC center name
MRC Clinical Sciences Centre, Imperial College London, Du Cane Road, London W12 0NN, UK
INSDC first public
2014-09-20T17:03:04Z
INSDC last update
2018-03-08T17:08:16Z
INSDC status
public
Submitter Id
E-MTAB-2237:rrp41.1_rna
broker name
ArrayExpress
common name
fission yeast
genotype
h? ade6-M21? leu1-32 ura4-D18 his3-D1 p81nmt1::kanMX6-rrp41
sample name
E-MTAB-2237:rrp41.1_rna
scientific_name
Schizosaccharomyces pombe
strain
FBY-1347