Abstract:
The difference between bitter and not bitter cassava based on the linamarase gene is
scientifically unknown. In order to know this difference, the scientific information is
necessary and can be conducted by isolation of total DNA and DNA amplification.
DNA amplification was done using two primer pairs (LK and LP) with different
annealing temperature (Ta). The average melting temperature (Tm) of LK and LP
primers were 52,3oC and 53,1oC, respectively. Ta values was determined by reducing
3°C or 5°C of Tm value. This study aimed to determine the annealing temperature of
the linamarase primer used in PCR. In this study, annealing temperature optimization
was carried out at 49,3oC and 47,3oC for LK primers and 50,1oC and 48,1oC for LP
primers. The template DNA used was total DNA from cassava (Manihot esculenta
Crantz.) genotype menggalo. The annealing temperature optimization of LK primer
produced a thick band with smear at both temperature (49,3oC and 47,3oC), while for
LP primer produced thick band without smear at 50,1oC, and with smear at 48,1oC.
Therefore, the primer used for amplification of linamarase gene was LP primer with
the annealing temperature was 50,1oC