2025-12-04
ENA first public
ERC000011
ENA-CHECKLIST
Berlin institute for medical systems biology
INSDC center name
public
INSDC status
ERS21154660
SRA accession
E-MTAB-14530:PHF3_dPHD.dTLD_br2
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
dPHD-dTLD
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
Homo sapiens
scientific_name
PHF3_dPHD.dTLD_br2
title