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Advances in Animal and Veterinary Sciences

Research Article
Adv. Anim. Vet. Sci. 1 (4S): 20 - 23. Special Issue-4 (Progress in Research on Viruses and Viral Diseases)
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Prasad Minakshi *, Koushlesh Ranjan, Pawan Kumar, Gaya Prasad
Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, India
*Corresponding author:

Polyacrylamide gel electrophoresis (PAGE) is a commonly used technique for analysis of RNA samples because of its low cost, ease of use and high sensitivity. Following one dimensional or two dimensional electrophoresis of RNA sample on a gel, the RNAs are typically visualized by some form of nucleic acid staining techniques. Of these commonly used methods employ use of intercalating fluorescent agents such as ethidium bromide (EB), SYBR green and SYBR gold under short wavelength UV Irradiation. Unfortunately these staining methods are potentially mutagenic and UV radiations are hazardous to both DNA samples and persons handling it. A novel method of RNA staining has been developed using silver ion sensitizer, Eriochrome Black t (EBT). Bluetongue viral RNA resolved in non–denaturing RNA polyacrylamide gel electrophoresis (RNA–PAGE) was used to develop the staining protocol. The method using Eriochrome Black t with silver staining (EBT–SS) has been found to be eight times more sensitive as compared to routine method used in the laboratory. In the routine method of silver staining procedure all the ten bands of bluetongue virus genome were visualized up to 0.625ng RNA sample per lane. However, in newly developed method, using EBT–SS the same was visualized up to 0.078ng RNA sample per lane. The silver ion sensitizer method could be applied for detection of RNA in various biological samples due to its higher sensitivity.

Key Words: Bluetongue virus, EBT, Silver staining, RNA–PAGE