organism: Homo sapiens
description: Protocols: Cell cultures were established from metastatic lesions surgically removed from cutaneous CM patients that did not underwent prior BRAFi therapy who were referred to the National Cancer Institute of Aviano (Italy), as previously described [1]. Cell cultures were grown in RPMI 1640 Medium, supplemented with 2mM L-glutamine (Sigma-Aldrich) and 10% heat-inactivated fetal calf serum (FCS, Lonza). The identity of paired Spry1KO clones and parental cells was confirmed by short tandem repeat profiling using the Power Plex 1.2 kit (Promega). Stock solutions of UO126 (Sigma), vemurafenib (PLX-4032, SeleckBio), trametinib (GSK1120212, SeleckBio) were prepared in cell-culture grade DMSO (Sigma) as per manufacturer's indications. The study was approved by the Internal Review Board of the Centro di Riferimento Oncologico, IRCCS-National Cancer Institute, Aviano, Italy (IRB number 07-2017). In details , indexed libraries were prepared using 1 ug of total RNA as starting material, with TruSeq Stranded Total RNA Sample Prep Kit (Illumina Inc.). Libraries were sequenced (paired-end,2x100 cycles) at a concentration of 8 pM/lane on the HiSeq 2500 platform (Illumina Inc). In details , indexed libraries were prepared using 1 ug of total RNA as starting material, with TruSeq Stranded Total RNA Sample Prep Kit (Illumina Inc.). Libraries were sequenced (paired-end,2x100 cycles) at a concentration of 8 pM/lane on the HiSeq 2500 platform (Illumina Inc).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-01-03T15:59:19Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-04-16T14:46:41Z
attributes.tag: External Id
attributes.value: SAMEA5568546
attributes.tag: INSDC center name
attributes.value: Laboratory of Molecular Medicine and Genomics
attributes.tag: INSDC first public
attributes.value: 2020-01-03T15:59:19Z
attributes.tag: INSDC last update
attributes.value: 2019-04-16T14:46:41Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7886:Sample 5
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: cutaneous melanoma
attributes.tag: genotype
attributes.value: BRAFV600E
attributes.tag: individual
attributes.value: 599
attributes.tag: organism part
attributes.value: skin
attributes.tag: sample name
attributes.value: E-MTAB-7886:Sample 5
attributes.tag: sampling site
attributes.value: neoplasm
attributes.tag: scientific_name
attributes.value: Homo sapiens