2025-12-04
ENA first public
ERC000011
ENA-CHECKLIST
Berlin institute for medical systems biology
INSDC center name
public
INSDC status
ERS21154668
SRA accession
E-MTAB-14530:PHF3_WT_br1
Submitter Id
ArrayExpress
broker name
HEK293
cell_line
not collected
collection date
human
common name
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 of 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 (21130 g) 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 30 min at 37°C and subsequently purified by phenol-chloroform extraction and ethanol precipitation. rRNA was depleted using RiboCop rRNA depletion kit (Human/Mouse/Rat) (Lexogen) and libraries were prepared using Corall total RNA Seq kit (Lexogen) according to the manufacturer’s instructions, starting with 600 ng total RNA input.
description
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
Homo sapiens
scientific_name
PHF3_WT_br1
title