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This study was carried out to investigate a usefulness of the microsatellite DNA markers for parentage verification of Thoroughbred (TB) horses. 9 TB horses samples were genotyped for nine international minimum standard markers (AHT4, 5, ASB2, HMS3, 6, 7, HTG4, 10, and VHL20), and the additional panel of four markers, ASB17, CA425, LEX33, and TKY321. This methods consisted of multiplexing PCR procedures, and it showed reasonable amplification of all PCR products. Genotyping was performed with an ABI 310 genetic analyzer. Foal Ⅰ was excluded according to principles of Mendelian genetics in AHT4 (H/K), ASB2 (Q/Q), HMS3 (I/P), HTG4 (M/O), HTG10 (K/R), VHL20 (M/P), ASB17 (F/N), LEX33 (M/O), and TKY321 (G/I) markers. Foal Ⅱ was excluded with markers AHT5 (K/M), ASB2 (M/N), HMS7 (N/N), HTG10 (K/K), VHL20 (I/I), ASB17 (F/F) and TKY321 (G/I). Foal Ⅲ was excluded with markers AHT4 (O/O), AHT5 (K/K), ASB2 (M/R), HMS6 (M/P), HMS7 (O/O), HTG10 (R/S), VHL20 (L/M), and ASB17 (N/O). These results suggest that the present DNA typing is so useful for parentage verification of TB horses.