2024-09-10
ENA first public
ERC000011
ENA-CHECKLIST
Medical Faculty Mannheim  University of Heidelberg Department of Anatomy and Developmental Biology
INSDC center name
public
INSDC status
ERS20914390
SRA accession
E-MTAB-14388:Sample 4
Submitter Id
ArrayExpress
broker name
PC-3
cell_line
not collected
collection date
human
common name
Protocols: PC-3 cells were grown in RPMI 1640 medium supplemented with 10% fetal calf serum (FCS) and 1.2% penicillin/streptomycin as described previously [10] (PAN-Biotech GmbH, Aidenbach, Germany). For RNA analysis, cells were grown to a subconfluence of 70-80%. The cells were treated with vehicle (0.005 M H2SO4) or 200 μmol of S-adenosylmethionine (SAM) (New England Biolabs, Ipswich, MA) for 120 hours. Under both conditions, the medium was changed every other day.   Total RNA was isolated from PC-3 cells using a NucleoSpin miRNA Kit (Macherey Nagel) according to the manufacturer’s instructions.   Following ligation of the RNA to modified 3 ́ and 5 ́ adaptors, the products were reverse transcribed (Super Script III; Thermo Fisher Scientific, Waltham, MA, USA), purified (SPRIselect reagent kit, Beckman Coulter)  and PCR amplified for 14 cycles (KAPA HiFi HotStart PCR Kit, KAPA Biosystems).Size selection was performed by polyacrylamide gel electrophoresis.
description
prostate cancer
disease
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
Homo sapiens
scientific_name
200 μM S-adenosylmethionine
stimulus
Sample 4
title