This paper reports on the ambipolar conduction for the λ-Deoxyribonucleic Acid (DNA) field effect transistor (FET) with 450, 400 and 250 base pair experimentally and theoretically. It was found that the drain current of the p-type DNA/Si FET increased as the ratio of the guanine-cytosine (GC) pair increased and that of the n-type DNA/Si FET decreased as the ratio of the adenine-thymine (AT) pair decreased, and the ratio of the GC pair and AT pair was controlled by the total number of the base pair. In addition, it was found that the hole conduction mechanism of the 400 bp DNA/Si FET was polaron hopping and its activation energy was 0.13eV. By considering the electron affinity of the adenine, thymine, guanine, and cytosine, the ambipolar characteristics of the DNA/Si FET was understood. The holes are injected to the guanine base for the negative gate voltage, and the electrons are injected to the adenine, thymine, and cytosine for the positive gate voltage.