DNA분석에 있어서 모세관 이동법과 진공압 이동법 및 전기 이동법의 비교

Abstract

In order to evaluate the efficiencies of various DNA blotting methods for routine clinical laboratory application of Southern blotting, we compared three blotting methods, i.e. capillary, vacuum, and electric. We used serially diluted concentrations of HindIII-digested lambda DNA for large molecular ranges (23.130-2.027 Kb), and 123 DNA ladder for small molecular ranges (0.123¬4.182 Kb). In capillary blotting, the efficiencies of DNA transfer both in large and small molecular ranges were good, but it took 18 hours to transfer DNA of 23 Kb in size, And the procedures were somewhat labor-intensive. In electric blotting, the efficiency of DNA transfer was good and blotting time was relatively short (2 hours). But the amount of blotting buffer consumption was as many as 5,000 ml and the cost of electric transfer apparatus was high. In vacuum blotting, the efficiencies of DNA transfer both in large and small molecular ranger were good, and blotting time was very short (only 1 hour). The amount of blotting buffer consumption was only 70 ml, and handling was very simple in vacuum blotting procedure. In conclusion, the vacuum blotting method is very rapid, simple, efficient and economic ； so we think this method is the best choice for routine clinical laboratory application of Southern blotting. Key Words : Southern blotting, Capillary transfer, Vacuum transfer, Electro-transfer.