Ekstrak glikogen temilok (Bactronophorus thoracites) sebagai Ko-presipitan asam deoksiribonukleat

Abstract

Temilok (Bactronophorus thoracites) is one of Teredinidae or family of wood-borer shellfish in fact it has a fairly high polysaccharide content, particularly interesting source of glycogen. The objectives of this experiment are to determine KOH concentration, percentation of cationic resin Amberlite IR-120 Na and its stirring time to extract the glycogen from temilok flesh based on Nicoletti and Baiocchi glycogen polysaccharide extraction patent method, and the ability of the extracted glycogen as co-precipitant in the precipitation of deoxyribonucleic acid (DNA) of human femur. Nitrogen content of extracted glycogen was measured by kjeldahl method. Nucleic acid content of extracted glycogen were spectrophotometrically measured at 260 nm, 280 nm and 320 nm. Extracted glycogen samples was characterized by determining the glucose content quantitatively using phenol-sulphate (phesul) method which measured spectrophotometrically at 490 nm . The treatment of 100g temilok flesh with 40% potassium hydroxide and heated to 100 oC decreased the nitrogen content of extracted glycogen to 250 ppm. Treated the solution with 12% resin cationic Amberlite IR-120 and stirring for 16 hours at room temperature yielded 10,19±2,23% glycogen characterized by the lowest nucleic acids content which is 0,07 mg/mL. In order to asses the ability of the glycogen extract as co-precipitant of low copy number DNA, 2% solution of glycogen extract were added until 20 μL per 800 μL solution of femur DNA and 40 μL sodium acetate. Results revealed that by adding the extracted glycogen solutions into the first step of DNA precipitation procedure, the yield of DNA were not significantly affected at the level of 0,05 though the yield of DNA decreased against blanko about 0,006 ng/μL of 0,0401 ng/μL.