An Efficient and Cost Effective Homogenisation Method for Isolating High Quality RNA from Fish Tissue Samples

Aquaculture sector has huge economic importance and provides jobs to millions of people besides providing nutritionally rich food to billions of people around the world. However, over the years the industry is challenged by various diseases that affect fish health and productivity, leading to economic losses and food security concerns. It is therefore important to understand the molecular basis of these diseases and develop new effective strategies. Transcriptomics is one of the powerful approaches in understanding the pathophysiology at molecular level. Extraction of the high quality RNA from the fish tissues has always remained a challenging task for transcriptomic analysis and tissue homogenisation is one of the initial and the most crucial step in RNA isolation. Here we report an easy, efficient and affordable method for homogenising fish tissues that yielded high quality RNA from fish tissues. The applicability of this method was evaluated for downstream applications including cDNA synthesis and RT-PCR analysis of housekeeping gene GAPDH. The experimental procedure consistently provided sufficient intact total RNA (n=30) and RIN number between 8.2-9.0.This homogenisation method would be highly useful for aquaculture researchers and help in understanding the transcriptomic changes in response to diseases or changed habitat.