Optimization of PCR conditions for amplification of heat shock protein 70 (HSP70) and 18s rRNA of cherax quadricarinatus

• Main Author: Athirah Abd Aziz
• Format: Undergraduate Final Project Report
• Published: 2016
• Online Access: http://discol.umk.edu.my/id/eprint/6348/

The polymerase chain reaction (PCR) conditions of heat shock proteins 70 (HSP70) and the housekeeping gene; 18S of crustacean’s species, the crayfish Cherax quadricarinatus were studied. In the PCR technique, PCR product was amplified by a series of polymerization. The specificity and yield of the products mainly depend on two parameters, which are the annealing temperature and the PCR cycle. The annealing temperature tested for 18S and HSP70 are ranging from 53°C to 65°C. The annealing temperature was optimized at 53°C for both genes and this temperature was used to optimize the PCR cycle. This temperature was resulted as evidenced by the products obtained in the gel electrophoresis. The PCR cycle for 18S and HSP70 gene was tested from 20-40 cycles and 24-44 cycles respectively. The best annealing temperature optimized for both genes were at 53°C and the best PCR cycles optimized for 18S was at cycle 23 and at cycle 39 for HSP70. The optimized PCR conditions can be need for future studies particularly in studying or investigating gene expression of HSP70 normalized with 18S rRNA.