degradement
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They include the variation of the degradement velocity of different sample, the inhibition product of PCR, the amount of DNA extraction, quality of DNA and the heteroplasmy of some tissue. Correct sequencing, comparison of the result of forward and reverse primers sequencing should be done to rule out wrong signals, and prevent contamination. In addition to gene sequencing, low grade heteroplasmy could be dealt by Denaturing high performance liquid chromatography. Multiplex Ligation-dependent Probe Amplification. The analysis of mitochondrial HV Ⅰ, HV Ⅱ sequencing is able to identify the maternal sanguinity, then it could assist in the human identification. It also complements the genomic STR system in distribution of population, the origin of human, and analysis for fossil.
分析粒线体DNA时必须考虑各类检体腐败DNA降解速度之不同、PCR的抑制物质、DNA的萃取量及品质、特定组织的异质现象、萤光序列分析图及正、反向引子定序结果比对、排除杂讯干扰、防治污染等,除以基因定序外,低程度的异质现象也可用Denaturing high performance liquid chromatography、Multiplex Ligation-dependent Probe Amplification等方法分析,粒线体HV Ⅰ、HV Ⅱ碱基序列的分析可鉴别一个人的母系血缘关系、辅助人类身分鉴定,对於族群分布、人类起源探讨、远古检体的分析可作为辅助STR系统的利器。