Silicalite-1 membrane was prepared on an outer surface of a tubular α-alumina support by a secondary growth method. Changes of defect amount and crystallinity during secondary growth were carefully observed. The defect-less well-crystallized silicalite-1 membrane was obtained after 7-days crystallization at 373 K. The silicalite-1 membrane became (h0l)-orientation with increasing membrane thickness, possibly because the orientation was attributable to “evolutionally selection”. The (h0l)-oriented silicalite-1 membrane showed high p-xylene separation performance for a xylene isomer mixture (o-/m-/p-xylene = 0.4/0.4/0.4 kPa). The p-xylene permeance through the membrane was 6.52 × 10−8 mol m−2 s−1 Pa−1 with separation factors αp/o, αp/m of more than 100 at 373 K. As a result of microscopic analysis, it was suggested that a very thin part in the vicinity of surface played as effective separation layer and could contribute to high permeation performance.