By utilizing a Mini-Primer strategy targeting the target site duplication (TSD) sequence of retrotransposons, insertion and null allele (INNUL) markers, which include short interspersed nuclear elements (SINEs), long interspersed nuclear elements (LINEs), and composite SVA retrotransposons (SINE/VNTR/Alu, where VNTR represents “variable number of tandem repeats” and Alu represents a type of primate specific SINE that has reached a copy number in excess of one million in the human genome), can be effectively used as markers for human identification and bio-ancestry studies regardless of the size of the inserted element. The size of the amplicons for INNULs and the difference between allelic states can be reduced substantially such that these markers have utility for analyzing high and low quality human DNA samples. Multiplexes including either 15 or 20 retrotransposable element (RE) markers plus Amelogenin for single tube amplification of DNA in four color detection were successfully designed. The multiplexes provided power of discrimination suitable for forensic and paternity analyses.
通过利用针对逆转录转座子目标位点重复(T
SD)序列的迷你rimer 策略,插入和无效等位
基因(INNUL)标记物,包括短穿插核元件(SINE)、长穿插核元件(LINE)和复合 SVA 逆转录转座子(SINE/VNTR/Alu、其中,VNTR 代表 "可变串联重复序列数",Alu 代表一种灵长类特有的 SINE,在人类
基因组中的拷贝数已超过一百万),无论插入元件的大小如何,都可有效地用作人类识别和
生物世系研究的标记。INNULs 扩增子的大小和等位
基因状态之间的差异可以大大缩小,因此这些标记物可用于分析高质量和低质量的人类 DNA 样本。我们成功地设计出了包括 15 或 20 个逆转录可逆元件(RE)标记和 Amelogenin 的多重试剂盒,用于在四色检测中对 DNA 进行单管扩增。这些多重标记的鉴别能力适用于法医和亲子鉴定分析。