Sequential extraction of RNA, DNA and protein from cultured cells of the same group

Core Tip

Core Tip: Sequential extraction of nucleic acids and proteins from cultured cells of the same group. 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. 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. 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. 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. 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. The quality of RNA, DNA and proteins isolated through sequential isolation protocol can be used for reverse transcription - polymerase chain reaction (PCR), PCR and western blot, respectively. The present procedure is not only easy, rapid and high efficient, but also economical and practical, especially for researchers in developing and underdeveloped countries.