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HeLa cell culture and small interfering RNA (siRNA)-mediated knockdown

MW Mengjun Wu
EK Evdoxia Karadoulama
ML Marta Lloret-Llinares
JR Jerome Olivier Rouviere
CV Christian Skov Vaagensø
MM Martin Moravec
BL Bingnan Li
JW Jingwen Wang
GW Guifen Wu
MG Maria Gockert
VP Vicent Pelechano
TJ Torben Heick Jensen
AS Albin Sandelin
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HeLa Kyoto cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. siRNA transfections (for SLIC-CAGE and TIF-seq) were carried out using lipofectamine 2000 (Invitrogen) according to the manufacturer's protocol. Cells were treated with 20 nM siRNA for 4 days, including a re-transfection 2 days after the initial transfection. siRNA sequences: siGFP: GACGUAAACGGCCACAAGUdTdT; siRRP40: CACGCACAGUACUAGGUCAdTdT; siZCCHC8: GGAAUGUACCUCAGGAUAAdTdT; siZFC3H1: GAUUAGAGUCCAUGAUUAAdTdT. RNA was extracted using TRIzol (Invitrogen) and treated with TURBO DNase (Invitrogen) following the manufacturer's instructions.

Copyright and License information:  ©2020 The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact moc.puo@snoissimrep.slanruoj

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