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Page No. : 91-95

ABSTRACT

A paper based device that integrates a silica fi lter disc as a DNA capture tool was successfully developed for low-cost, rapid and automated DNA extraction from chicken, goat and pig meat samples. The device was constructed by sandwiching a piece of silica pallet by two layers of nitrocellulose membranes and for absorption of digested protein and debris a cellulose paper was used. The basic principle used to extract genomic DNA using this device was solid phase approach. The trapping of DNA was done on solid phase silica based material. Digestion regime of 25 mg of minced muscle tissue sample was optimized for appropriate time-temperature combination and Proteinase K concentration. The developed extraction procedure used chemicals provided with DNeasy blood and tissue kit (Qiagen, Germany). The quality and purity of extracted DNA were assessed by Nanodrop spectrophotometer reading. The quality of extracted DNA was also visually checked bygel documentation system. For the appropriate digestion of 25 mg of muscle tissue samples, incubation time of 120 min at 56°C was found optimal. The concentrations of extracted DNAs were found in ranges of 15 to40 ng/μl and OD260:280 ranged between 1.6 and 1.9. These values indicated that extracted DNA using silica based filter paper assembly was good enough to carry out nucleic acid test (NAT).

Keywords: Genomic DNA, paper assembly, proteinase K, silica pallet