Avanços na Pesquisa em Ciências Aplicadas Acesso livre

Abstrato

Development of Midi-Plasmid Isolation Kit and Cost Optimization of DNA Molecular Weight Ladder: Economy of Laboratory vs. Commercial Products

Atul Kumar, Sachidanand Singh, Akash Tomar

Major impediments to biotechnology research are the excessive operational expenses involved in each set of experimental procedures not only due to use of high purity chemicals and enzymes but also various molecular biology Kits and other commercial products. This is a work which envisages to establish primary molecular biology protocols in lab to produce economical kits and products which will not only bring down the operational expenses, but also aid in the laboratory courses of the college. Preparation of competent cells of various E. coli strains is a mandatory requirement for transformation and cloning. The plasmid isolation protocol used for this purpose was midi preparation. Consequently, Plasmid Transformation, Restriction and a low cost molecular weight composite ladder (pBR322/ Hinf I+BamH I) has been prepared. The cost of ladder production could be reduced further by lowering the concentration of Restriction Enzyme and/ or increasing the incubation time for restriction digestion. Finally the cost of commercially available kits and ladder are compared to obtain the effective economy achieved in laboratory. These attempts will be beneficial as these standardized, economically efficient protocols/ products for competent cell preparation, plasmid isolation and molecular weight DNA ladder generation could be used for routine Laboratory experiments and embedded, even into critical experimental procedures.