The present invention relates to a method for preparation of a filer containing a DNA fraction and a filler prepared thereby. Despite low contents of the components in the filler, the preparation method of the present invention can secure viscoelasticity in the filler as a physical support and has a high efficiency due to the advantage of preparation processes thereof. The filler prepared through the filler preparation method of the present invention is superb in terms of skin thickness improvement, skin wrinkle reduction effect, and collagen formation ability and causes low pain during injection, giving high satisfaction to patients and operators.