PCR optimization for amplicafication of anti-lipopolysaccharide factor of redclaw (Cherax quadricarinatus)

This study presents a serial optimization of polymerase chain reaction (PCR) for amplification of Anti lipopolysaccharide factor (Alpf) gene fiom the redclaw crayfish, Cherax quadricarinatus. The redclaw crayfish was chosen because of lack of research done about the crayfish for immune related genes. These types of genes are important for the improvement of crayfish industry around the country. By Using PCR process, we can determine the optimized PCR condition of the specified genes, which is the first step on researching the gene against crayfish plague. This research investigates several parameters in
PCR optimization, including annealing temperature of the primers, quantity of the template and concentration of the MgCl₂. In the end, this will help researcher find ways to improve crayfish industry quality. Through a serial optimization, a band at desired size was obtained.
However, it is suggested to optimize the concentration of MgCl₂ to get a discrete single band. This work provides a preliminary reference to amplify the Alpf from redclaw crayfish specifically, hence to be applied for gene expression study in the fixture.