Volume: 3, 2024
3rd International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland:
ENVIRONMENT – PLANT – ANIMAL – PRODUCT
Abstract number: A010
DOI: https://doi.org/10.24326/ICDSUPL3.A010
Published online: 24 April 2024
ICDSUPL, 3, A010 (2024)
Comparison of column-based and organic GTPC methods of RNA isolation from pig liver
Paweł Grychnik1*, Kaja Ziołkowska1, Angelika Tkaczyk-Wlizło1, Krzysztof Kowal1, Brygida Ślaska1
1 Department of General and Molecular Genetics, Institute of Biological Basis of Animal Production, Faculty of Animal Sciences and Bioeconomy, University of Life Sciences in Lublin, 13 Akademicka, 20-950 Lublin, Poland
* Corresponding author: pawel.grychnik@up.lublin.pl
Abstract
RNA is a key component of cells and plays an important role in protein synthesis. There are many methods of RNA isolation from various types of tissues, including the extraction method using guanidinium thiocyanate, phenol, and chloroform (GTPC) and the extraction method using a silica matrix (SM). In the GTPC method, RNA is separated from DNA and proteins by centrifugation, and then precipitated with isopropanol. In the SM method, RNA is adsorbed onto a silica matrix, and contaminants are removed with a wash buffer. The purified RNA is eluted with DEPC-treated water or commercial RNase-free water. The research material used in the work are fragments of the liver of 5-6 week old piglets. The choice of pig liver is due to its similarity to the human liver, availability, and usefulness in further research. The aim of this work is to evaluate the efficiency and purity of the obtained RNA isolates in both of these methods. We want to determine which of these methods is more suitable for the isolation of RNA from pig liver. As part of this methodological development, the efficiency and purity of RNA isolates obtained using a column-based method, using a Qiagen kit, and a traditional isolation method based on TRIzol were compared. The method using TRIzol allows obtaining several to several times more ng RNA than in the case of the column method, with a similar purity A260/280 for both methods. It should also be noted that the method with TRIzol is longer to perform and TRIzol itself is toxic and improper isolation can harm health, while the column method is a shorter method but less often correctly comes out. This comparison provides valuable information about the effectiveness of these methods and can help in future research on gene expression.
The study was funded by NCN – OPUS-22 LAP, project number: 2021/43/I/NZ9/02612B.
Keywords: isolation, RNA, pig, liver
How to cite
P. Grychnik, K. Ziołkowska, A. Tkaczyk-Wlizło, K. Kowal, B. Ślaska, 2024. Comparison of column-based and organic GTPC methods of RNA isolation from pig liver. In: 3rd International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland: Environment – Plant – Animal – Product. https://doi.org/10.24326/ICDSUPL3.A010