It is difficult to differentiate between type 9.3 and 10 at the TH01 locus. Therefore, we designed three primers including allele-specific primer for the type 9.3. Our primer sets gave shorter fragments ranging from 62 to 81 bp than those in other reports, allowing each allelic band to be resolved by 10% non-denaturing polyacrylamide gel electrophoresis and stained with SYBR Green I. TH01 typing was successfully applied to human remains such as sera, whole bloods and blood stains made about 80 years ago without extracting DNA from them. The simple and fast methods for TH01 typing as demonstrated herein will be useful in the field of forensic practice.