OPTIMALISASI KONSENTRASI POLIAKRILAMID UNTUK DETEKSI MUTASI GEN PADA METODE POLYMERASE CHAIN REACTIONSINGLE STRAND CONFORMATION POLYMORPHISM (PCR-SSCP) DAN UJI PELABELAN ISOTOPIKNYA

Single strand conformation polymorphism (SSCP) method is one of further analysis methods in genetics that use
polymerase chain reaction (PCR) product for fast detection of genetic mutation based on the alteration of
nucleic acid migration on non denatured polyacrylamide gel. This method is also widely used in biodiversity
study. Aim of the research was to get an optimal concentration of acrylamide so that the best visualization of deoxyribonucleic acid (DNA) was obtained. Gel was made in certain composition by varying the percentage of
acrylamide-bis-acrylamide monomer between 15 to 30% (percentage of volume) and followed by electrophoresis
of single strand DNA for a certain time and visualized. The additional of glycerol was also examined at one
concentration. Results showed that the best concentration was found at 25% of acrylamide without glycerol. Test
on the isotopic labeling of DNA with 0.1 µCi of P-32-dCTP and autoradiography showed a better quality in
visualization compared to that of non-labeled one