4_CTR_BC_6
Homo sapiens
Five hundred thousand HFFF2 cultured for seven days Protocols: Human Foetal Foreskin Fibroblasts 2 (HFFF2) are cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% Foetal Bovine Serum and 292 ug/ml L-Glutamine. They have been treated with 2ng/ml TGF-beta-1 to induce myofibroblast differentiation or with 10Gy gamma irradiation to induce senescence. We isolated RNA 7 days upon this treatment changing medium 3 days bbefore the RNA extraction using RNAeasy minikit from Qiagen The kit we used is \Truseq Stranded mRNA\ sample prep kit from Illumina. Basic principle for stranded RNA library preparation is that dUTP is incorporated in the second strand synthesis of the cDNA. After adapter ligation, the dUTP strand is degraded by Uricil-DNA-Glycosylase (UDG) and the remaining strand is then amplified to make the Illumina library. However, Illumina uses polymerase I which is intolerant to dUTP, thus it just does not amplify the strand with dUTP incorporated. More details can be found in the Illumina's Guide on page 2 in the link down here: http://support.illumina.com/content/dam/illumina-support/documents/documentation/chemistry_documentation/samplepreps_truseq/truseqstrandedmrna/truseq-stranded-mrna-sample-prep-guide-15031047-e.pdf
ENA-FIRST-PUBLIC
2017-06-01T17:01:55Z
ENA-LAST-UPDATE
2018-03-08T22:40:15Z
External Id
SAMEA3212974
INSDC center name
University of Southampton
INSDC first public
2017-06-01T17:01:55Z
INSDC last update
2018-03-08T22:40:15Z
INSDC status
public
Submitter Id
E-MTAB-3101:4_CTR_BC_6
broker name
ArrayExpress
cell line
HFFF2
cell type
foetal foreskin fibroblast
common name
human
growth condition
DMEM supplemented 10% FBS and 10% CO2
sample name
E-MTAB-3101:4_CTR_BC_6
scientific_name
Homo sapiens