Ble-3
Homo sapiens
Protocols: Samples were lysed on plate using RNeasy kit (Qiagen) Lysis buffer. RNA was extracted using RNeasy kit (Qiagen) using manufacturer protocol. DNA was removed in column treatment and in-solution with DNAse I. Individual libraries were generated from 500ng of each total RNA sample using the QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) for Illumina (Lexogen Inc, #015). Libraries were prepared according to the QuantSeq 3' mRNA-seq Library Prep Kit (FWD) protocol. The QuantSeq protocol generates only one fragment per transcript, resulting in extremely accurate gene expression values, and the sequences obtained are close to the 3' end of the transcripts. Library generation was initiated by oligo-dT priming. The primer already contains Illumina-compatible linker sequences. After first strand synthesis, the RNA was removed and second strand synthesis initiated by random priming and a DNA polymerase. With the QuantSeq FWD kit the Read 1 linker sequence is contained in the second strand synthesis primer so NGS reads are generated towards the poly (A) tail and directly correspond to the mRNA sequence. Second strand synthesis was followed by a magnetic bead-based purification step before cDNA libraries were amplified for 13 cycles introducing the sequences required for cluster generation and index sequences to allow multiplexing of sequencing libraries. After amplification a final bead-purification removed any excess primers or adapterdimers that may interfere with sequencing.
INSDC center name
ED
Submitter Id
E-MTAB-13292:Ble-3
broker name
ArrayExpress
cell_line
HEK293
cell_type
epithelial cell
collection date
not collected
common name
human
developmental stage
embryo
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
kidney
phenotype
wild type
sample name
E-MTAB-13292:Ble-3
scientific_name
Homo sapiens