Cui YY. Sequential extraction of RNA, DNA and protein from cultured cells of the same group. World J Methodol 2023; 13(5): 484-491 [PMID: 38229947 DOI: 10.5662/wjm.v13.i5.484]
Corresponding Author of This Article
Ying-Yu Cui, PhD, Associate Professor, Teacher, Department of Cell Biology, Institute of Medical Genetics, Key Laboratory of Arrhythmias of the Ministry of Education of China, Tongji University School of Medicine, No. 500, Zhennan Road Putuo District, Shanghai 200331, China. yycui@tongji.edu.cn
Research Domain of This Article
Biochemical Research Methods
Article-Type of This Article
Basic Study
Open-Access Policy of This Article
This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
World J Methodol. Dec 20, 2023; 13(5): 484-491 Published online Dec 20, 2023. doi: 10.5662/wjm.v13.i5.484
Sequential extraction of RNA, DNA and protein from cultured cells of the same group
Ying-Yu Cui
Ying-Yu Cui, Department of Cell Biology, Institute of Medical Genetics, Key Laboratory of Arrhythmias of the Ministry of Education of China, Tongji University School of Medicine, Shanghai 200331, China
Author contributions: Cui YY designed and performed the research, and analyzed the data and wrote the manuscript.
Supported bythe Postdoctoral Science Foundation of China, No.2005038300; and the National Natural Science Foundation of China, No. 30671028.
Institutional review board statement: No human and/or animal subjects was involved in the present study.
Institutional animal care and use committee statement: This manuscript does not involve in animal model.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: This manuscript does not involve in animal model.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Ying-Yu Cui, PhD, Associate Professor, Teacher, Department of Cell Biology, Institute of Medical Genetics, Key Laboratory of Arrhythmias of the Ministry of Education of China, Tongji University School of Medicine, No. 500, Zhennan Road Putuo District, Shanghai 200331, China. yycui@tongji.edu.cn
Received: July 18, 2023 Peer-review started: July 18, 2023 First decision: September 13, 2023 Revised: September 25, 2023 Accepted: October 16, 2023 Article in press: October 16, 2023 Published online: December 20, 2023 Processing time: 155 Days and 7.1 Hours
ARTICLE HIGHLIGHTS
Research background
Life is a way of material (mainly protein and nucleic acid) movement and health lies in movement. Cell is the most fundamental structural and functional unit of life, therefore, the effective isolation of nucleic acids and proteins from cells is the foundation and prerequisite for revealing the mysteries of life. However, during laboratory routine for isolation of nucleic acids and proteins, cell samples are often from different culture dishes, usually leading to inevitable experimental errors and sometimes poor repeatability.
Research motivation
The present research tries to explore the possibility to simultaneously isolate nucleic acids and proteins from the same sample, while reducing experimental errors and ensuring consistency during experimentation.
Research objectives
The present study established a selective protocol for sequential isolation of RNA, DNA and proteins from the same cells with the characteristics of easy operation, rapid extraction and high efficiency.
Research methods
RNAzol reagent was used for the sequential isolation of RNA, DNA, and proteins from the same cultured HepG2 cells, resulting in a novel protocol containing four steps.
Research results
A protocol for sequential isolation of RNA, DNA and proteins was established and the procedure takes as few as 3-4 d from the start to quality verification and is highly efficient.
Research conclusions
The quality of RNA, DNA and proteins isolated through sequential isolation protocol can be used for reverse transcription (RT) - polymerase chain reaction (PCR), PCR and western blot, respectively.
Research perspectives
The present procedure is not only easy, rapid and high efficient, but also economical and practical, especially for researchers in developing and underdeveloped countries.
