Forensic single cell separation testing technology is one of the most effective ways for trace and mixture DNA sample testing. Traditional single cell separation testing technology includes single cell isolation and lysis, PCR amplification and data analysis. This article details the principle of traditional single-cell separation and its application in case testing, meanwhile places the focus on prospect of single-cell sequencing (SCS) technology into forensic application. The research on SCS has shown tremendous growth over the past five years, and impacted many diverse areas of biological research. However, the study in forensic genetics is as yet waiting for developing. SCS is a combination of the single cell isolation, similar to the traditional single cell separation, with whole-genome amplification and next generation sequencing. In this review, the whole genome amplification and next generation sequencing (NGS) were summarized together with the advantages and disadvantages of the current technologies analyzed. Until now, there are some commercial kits of whole-genome amplification for biological research and ones of next generation sequencing for forensic application. Although the SCS technology is still relatively new, it has been being developed rapidly and will provide new means for forensic testing, especially for trace and mixture sample testing. The grounds root at the quantity-enormous parallel sequencing of NGS that is relied with SCS. This implementation can make one single sample tested hundreds of times so that such repetitious verification will definitely produce more complete and accurate information. Besides, next generation sequencing can bring the minor component in mixed sample into its STR types, even leading the mixture containing substance of three or more persons to evidential results, therefore exposing various genetic markers like STR, SNP from different individuals in just one single run of detection. Additionally, the real-time single molecule’s sequencing by the third generation sequencing will set a progressive aspiration for forensic genetics with the direct reading DNA sequence.%法医单细胞分离检验技术是微量混合 DNA 样本检验最有效的方法之一。常规的单细胞分离检验技术包括单细胞分离、单细胞裂解、PCR 扩增、数据分析。本文将详细介绍单细胞分离检验技术原理和应用进展，同时展望了单细胞测序在法医 DNA 的应用前景。近5年来，单细胞分离、全基因组扩增和二代测序技术的结合而产生的单细胞测序技术，将为法医遗传学检测提供一种全新的技术和思路，尤其为微量混合生物样品检验提供全新的技术手段。
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