OPTIMASI SUHU ANNEALING PRIMER UNTUK AMPLIFIKASI DNA GEN COX3 MITOKONDRIA PADA Kryptopterus apogon (Bleeker 1851)
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Date
2016-01-26
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Abstract
The genetic information of COX3 mitochondria gene from Kryptopterus apogon is still unknown. Isolating and amplifying of total DNA is the first step before having the genetic information. In this study, the DNA amplification was performed using a pair of primer i.e. COX3F (F1) and COX3R (R1) with different melting temperature (Tm). The Tm of F1 and R1 primers are 54,3°C and 52,9°C, respectively. This study aimed to determine the annealing temperature of the COX3 mitochondria gene primers which have different melting temperature. The annealing temperature can be determined based on the average Tm of both primer and reduce it to 2°C or 5°C. Optimization PCR with the average Tm reduce for 2ºC (51,6ºC) during annealing produce thicker band compared to average Tm reduce for 5ºC (48,6°C). Therefore the suitable annealing temperature for F1 and R1 COX3 mitochondria primers was 51,6°C.
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Keywords
Annealing temperature, COX3 mitochondria gene, Kryptopterus apogon, melting temperature (Tm), PCR optimation