Assisted by the mold preinstalled an alumina tube into seed segment, the influence of the original primary dendrite spacing of seed on the formation of stray grains at melt-back region was investigated during Ni-based single crystal casting using seeding method. The results showed that the interface reaction between the seed and mold as well as the formation of stray grains at the surface of seed were avoid using an alumina tube with surface roughness of 0.35μm as the mold inner wall. As the original primary dendrite spacing of the seed decreased to less than 201μm, the morphology of un-melted solid phase in semi-solid zone changed from isolated dendrite stem to the complex network, resulting in the inhibition of the formation of stray grains inside of the seed. The seed with original primary dendrite spacing of 201μm was also successfully re-used to fabricate single crystal casting.