Abstract: Objective To establish optimal amplification conditions with the application of 0.2 mL test tube in single cell separation and inspection. Methods Oral epithelium cell suspension was prepared. Five or ten cells were collected with 0.2 mL test tube. Then DNA were amplified with Identifiler?誖Plus kit in three different conditions in which the proteinase K addition, gold enzyme concentration in PCR reaction, and PCR reaction cycles were adjusted separately. Finally the detection rate, allelic dropout rate and artificial alleles were compared among the groups. Results In these 3 different conditions： addition of proteinase K, addition of 0.4 μL gold enzyme in PCR reaction, and use of 32 cycles, the detection rate was higher and allelic dropout rate was lower than the other conditions. Conclusion In single cell separation and inspection, the usage of 0.2 mL test tube could be an effective supplement to chip-low volume amplification.

Key words: forensic genetics, nucleic acid amplification techniques, single cell