Since standard routine protocols yield unacceptable results in pancrease, we have designed a simple method for RNA Doxorubicin mouse extraction by comparing different protocols. Methods: We obtained 20-30 mg pancreatic tissues in less than 2 min from 30 rats. Several methods were performed to extract RNA from pancreatic tissue and evaluate its integrity. All methods were performed three times to obtain reproducible results.

Results: Immersing pancreatic tissue Inhibitors,research,lifescience,medical in RNA-later for 24 h at -80ºC yielded high quality RNA by using the TriPure reagent which was comparable to the commercial RNeasy Micro Kit. The quality of RNA was evaluated by spectrophotometer, electrophoresis and RT-PCR. We separated intact 28S and 18S ribosomal RNA (rRNA) when our procedure was compared with the RNeasy Micro Kit. Finally, full length of the actin gene was amplified by RT-PCR. Conclusion: We designed a simple, Inhibitors,research,lifescience,medical fast, cost-effective method for complete RNA extraction from the least amount of quantitatively intact pancreatic tissue. Keywords: Extraction, RNA, Pancreas, Autolysis Introduction Information of a structural gene is usually transcripted to a functional product by gene expression. Recent studies have focused on RNA analysis as a gene

expression tool in cells to detect differential gene expression between two conditions. Different methods have been presented for Inhibitors,research,lifescience,medical extracting nucleic acids such as guanidinium thiocyanate followed Inhibitors,research,lifescience,medical by phenol-chloroform extraction, chromatography by cellulose, extraction using silica matrices, magnetic bead based nucleic acid purification, and anion-exchange.1,2

Accurate detection of gene expression is influenced by status of the RNA that is isolated from tissues. The quality of isolated RNA should be checked prior to its use in subsequent tests and studies. The purity and quality of the isolated RNA is a vital step in RNA dependent assays. Performing complementary molecular tests with low-quality RNA Inhibitors,research,lifescience,medical may compromise the results of downstream applications which are often labor-intensive, time consuming, and highly expensive. Researchers need high quality RNA for molecular biological tests that have various diagnostic applications Chlormezanone such as quantitative RT-PCR, micro-arrays, ribonuclease protection assay, northern blot analysis, RNA mapping, and cDNA library construction.3,4 The quality of purified RNA from tissues and cells is variable. Often, after extraction, RNA is rather unstable over a long storage time. Long mRNA fragments up to 10 kb are especially sensitive to degradation.5,6 Researchers must consider various factors that affect the quality of purified RNA. Purified RNA must not be contaminated with RNases, proteins, genomic DNA, and enzymatic inhibitors. Additionally, the UV absorption ratio (260/280) of total RNA should be between 1.8-2.0 and RNA should have a minimal degree of fragmentation during electrophoresis.