Quantitative reverse transcription-PCR (qRT-PCR)

Xiaobin Wang; Xiaoyu Liu; Huihan Wang

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Quantitative reverse transcription-PCR (qRT-PCR)

XW Xiaobin Wang

XL Xiaoyu Liu

HW Huihan Wang

This method is extracted from research article: Cancer Gene Ther, Jun 2022

Combination regimen of granulocyte colony-stimulating factor and recombinant human thrombopoietin improves the curative effect on elderly patients with leukemia through inducing pyroptosis and ferroptosis of leukemia cells

DOI: 10.1038/s41417-022-00497-8

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Total RNAs from HL-60 and KG1a cells were extracted using TRIZOL (Invitrogen, USA). The first-strand cDNA was made by PrimeScript II 1st Strand cDNA Synthesis Kit (TaKaRa Biotechnology, Dalian, Liaoning, China) according to the manufacturer’s instructions. The amount of target RNA was normalized to the amount of internal control (GAPDH) and the results were given by 2^−△△Ct relative to the control sample. The qRT-PCR was performed by SYBR Green (Takara Biotechnology, China). The primer sequence was as follows: GPX4 forward: 5′-TCAGCAAGATCTGCGTGAAC-3′, reverse: 5′-GGGGCAGGTCCTTCTCTATC-3′; EP300 forward: 5′-CCTGAGTAGGGGCAACAAGAAGA-3′, reverse: 5′-ATGAGGCGGATCACAAAGAAGAC-3′; GAPDH forward: 5′-AACGGATTTGGTCGTATTGGG-3′, reverse: 5′-CCTGGAAGATGGTGATGGGAT-3′.

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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