Analysis of Laboratory skills and Chemical Preparation of Student of Packaging and Material Technology Program, Faculty of Agro-Industry

Abstract

Ethidium bromide is commonly used to stain DNA in molecular biology laboratories. Because this chemical can efficiently intercalate with nucleic but ethidium bromide is a mutagenic compound. Despite its advantages, its reputation as a mutagen activated the development of many alternative agents for safety reasons.  Prior to use these alternative chemicals, however, the comparative study of DNA staining effectiveness between the alternatives and Ethidium bromide is needed to be quantified.  In this study there were two commercial alternatives, GelRed® and SYBR® Safe, used to compare the effectiveness of DNA staining with Ethidium bromide. The brightness of DNA bands that appeared in agarose gel after electrophoresis were compared between the treatments of different staining agents, 15 wells each.  The gels were then taken photos under UV light, and the mean gray value of each DNA band was using Adobe Photoshop software to represent the brightness of each band.  The result indicated that the DNA staining effectiveness of the agents was different (p<0.0001).  Both alternative agents showed brighter DNA band under UV light rather than Ethidium bromide (p<0.0001 for both pairs) but they show no difference from each other (p=0.5570).  This study suggested that GelRed® and SYBR® Safe can be effectively used instead of ethidium bromide to reduce toxic waste, as well as to promote safety standards in the laboratory according to the standard of the Enhancement of safety Practice of Research Laboratory in Thailand (ESPReL).