HIGH-YIELDING TOTAL RNA ISOLATION OF CHICKEN LIVER USING LITHIUM CHLORIDE-BASED PURIFICATION METHOD FOR POLYSACCHARIDE-RICH TISSUES

Abstract

The fidelity of transcriptome profiling and the subsequent construction of reliable libraries for gene expression analysis are contingent upon the quality of isolated RNA. While high-yielding extraction protocols for plant tissues are well established, current methods can be less efficient for animal tissues, particularly those polysaccharide- and metabolite-rich samples, such as the liver. Commercial kits often provide solutions but are costly, limiting accessibility in resource-constrained laboratories. This study reports an optimized total RNA isolation protocol using lithium chloride (LiCl)-based purification to address these limitations. The method effectively eliminated glycogen co-precipitation and contamination through stepwise purification. Comparative analysis demonstrated that RNA purity (2.15 OD ratio) and yield (150.45 ng/µL) were statistically comparable to conventional and spin-column methods (p = 0.011 and 0.004, respectively). The RNA isolates demonstrated suitability for downstream molecular applications, as confirmed by successful cDNA synthesis and PCR amplification of the β-actin housekeeping gene. By combining efficiency, reproducibility, and affordability, this LiCl-based protocol offers a broadly applicable approach for RNA isolation.