RNA Extraction from Animal and Human's Cancerous Tissues: Does Tissue Matter?

Document Type : Original Article

Authors
Ali Akbar Samadani 1
Novin Nikbakhsh 2
Sadegh Fattahi 1
Roghayeh Pourbagher 1
Seyyed Mohsen Aghajanpour Mir 1
Narges Mousavi Kani 1
Zeinab Abedian 1
Haleh Akhavan-Niaki 1
1 Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Iran
2 Department of Surgery, Babol University of Medical Sciences, Babol, Iran
Abstract
The reliability of gene expression profiling, based technologies and methods to find transcriptional differences
representative of the original samples is influenced by the quality of the extracted RNA. Hence, RNA extraction
is the first step to investigate the gene expression and its function. Consequently, the quality of extracted RNA is
really significant. Correspondingly, this research was accomplished to optimize the RNA extraction methods and
compare the amounts of tissue or quality of tissue. Relatively, the cancerous tissue of human stomach in fresh
and frozen conditions and also the mouse fresh tissue were studied. Some factors like the amount of samples,
efficacy differences of diverse extraction buffers (TriPure, Trizol) and also the efficacy of b-mercaptoethanol
were compared and investigated. The results indicated that the less amount (1-2 mg) compared to other amounts
(2-5 mg, 5-15 mg) yielded the best quality and the RNA bands (5S, 18S, 28S) were observed perfectly.
Relatively, comparing and measuring some kinds of buffers (Trizol, TriPure) indicated no difference in RNA
extraction quality. The last investigated factor was the effect of b- mercaptoethanol which was used along with
TriPure to remove the RNAse. Conclusively, no effective impression was observed.
Keywords
RNA extraction
frozen tissue
cancerous tissues