Homo sapiens
Protocols: Cells from a 90% confluent 10 cm dish were harvested and counted, 8 million cells were mixed with 2 million Drosophila S2 cells as a spike-in control. Cell pellets from one well on a 6-well plate were resuspended in 1 mL TRI reagent (Sigma) and incubated for 5 min at room temperature. 200 µL chloroform (Applichem) was added and the lysate was vortexed and centrifuged at max. speed (21130g) for 15 min at 4°C. The aqueous layer was transferred to a new tube and precipitated with 0.5 mL isopropanol. The RNA pellet was isolated by centrifugation for 30 min at 4°C, washed with 1 mL 75% ethanol, re-centrifuged for 10 min, dried and resuspended in 70 µL RNase free water. 20 µg RNA was treated with 40 U DNaseI (Roche) for 30min at 37°C and subsequently purified by phenol-chloroform extraction and ethanol precipitation. rRNA was depleted using NEBNext rRNA depletion kit v2 (Human/Mouse/Rat) (NEB) and libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB) according to the manufacturer's instructions, starting with 600 ng total RNA input.
ENA-FIRST-PUBLIC
2023-11-16T19:00:05Z
ENA-LAST-UPDATE
2023-11-16T19:00:05Z
External Id
SAMEA114625728
INSDC center name
BIMSB
INSDC first public
2023-11-16T19:00:05Z
INSDC last update
2023-11-16T19:00:05Z
INSDC status
public
Submitter Id
E-MTAB-12782_2:RNA_HEK_PHF3_WT_br2
broker name
ArrayExpress
cell_line
HEK293
collection date
not collected
common name
human
developmental stage
embryo
genotype
PHF3_WT
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
kidney
sample name
E-MTAB-12782_2:RNA_HEK_PHF3_WT_br2
scientific_name
Homo sapiens