Patent ID: 9255276
Filing Date: 2016-02-09
CPC Classification: A61N,C12N

Claim Text:
1. A method of transferring an exogenous gene into a green alga cell, the method comprising performing electroporation using multiple square-wave pulses by the following steps: applying a square-wave electric pulse that satisfies a condition as defined in the following item (A) to a solution one time, two times or more so that the square-wave electric pulse has a total electric energy of 1.3 to 4.9 J/100 μL, the solution comprising the green alga cell with cell-wall, and a nucleic acid molecule; applying a square-wave electric pulse that satisfies conditions as defined in the following items (B1) and (B2) to the solution at least two times; and applying a square-wave electric pulse that satisfies conditions as defined in the following items (C1) to (C3) to the solution at least two times: (A) a square-wave electric pulse having a voltage per pulse of 750 V/cm or more; (B1) a square-wave electric pulse having a voltage per pulse of 150 V/cm or less; (B2) a square-wave electric pulse having an electric energy per pulse of 0.02 to 0.6 J/100 μL; (C1) a square-wave electric pulse opposite in polarity to the electric pulse that satisfies the conditions as defined in the items (B1) and (B2); (C2) a square-wave electric pulse having a voltage per pulse of 150 V/cm or less; and (C3) a square-wave electric pulse having an electric energy per pulse of 0.02 to 0.6 J/100 μL.