Efisiensi Transformasi Plasmid Pta7002-Atrkd4 Pada Escherichia Coli Bl21(De3) Dengan Metode Kejutan Panas

Tanio, Eunike Priscilla (2017) Efisiensi Transformasi Plasmid Pta7002-Atrkd4 Pada Escherichia Coli Bl21(De3) Dengan Metode Kejutan Panas. .. pp. 1-13. ISSN .

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Abstract

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

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