Plasmid transformation into a host cell is one of an important method in
gene cloning and usually used heat-shock metho, which can be affected by heatshock
time and temperature. This research main goal is to know how these two
factors affected heat-shock transformation of pTA7002-AtRKD4 using E. coli
BL21(DE3) as a host cell. This research have the following step, such as
pTA7002-AtRKD4 isolation from Agrobacterium tumefaciens EHA105, plasmid
purity and quantity test, detection of HPT and AtRKD4 gene, competent cell,
plasmid heat-shock transformation using randomized factorial design with two
variables (heat-shock temperature (42oC, 44oC, and 46oC) and heat-shock time
(30s, 60s, 90s, 120s, and 240s), triplicate), bacterial selection with kanamycin and
hygromycin, and determination and statistical analysis of pTA7002-AtRKD4
transformation efficiencies. Based on transformation efficiency calculation, the
optimum heat-shock transformation is 44oC for 30 seconds