Dugan Kerri A, Lawrence Helen S, Hares Douglas R, Fisher Constance L, Budowle Bruce
Laboratory Division, Federal Bureau of Investigation, Quantico, VA 22135, USA.
J Forensic Sci. 2002 Jul;47(4):811-8.
Mitochondrial DNA (mtDNA) analysis of forensic samples typically is performed when the quantity and quality of DNA are insufficient for nuclear DNA analysis or when maternal relatives may be the only reference source. Many of the steps required in the analytical process are both lengthy and labor intensive. Therefore, improvements in the process that reduce labor without compromising the quality of the data are desirable. The current procedure requires purification of the amplicons by centrifugal filtration after PCR and prior to cycle sequencing. Because this method requires several manipulations to perform, alternate cleanup procedures were investigated. These include the use of 1) Qiagen QlAquick PCR Purification columns, 2) Concert Rapid PCR Purification columns, and 3) ExoSAP-IT reagent. When the yield of purified amplicons, quality of the sequence profile, and ease of assay were evaluated, the use of ExoSAP-IT reagent for post-amplification purification was chosen to replace the filtration method.
法医样本的线粒体DNA（mtDNA）分析通常在DNA的数量和质量不足以进行核DNA分析时进行，或者当母系亲属可能是唯一的参考来源时进行。分析过程中所需的许多步骤既冗长又耗费人力。因此，需要改进该过程，在不影响数据质量的情况下减少劳动力。当前的程序要求在PCR后和循环测序前通过离心过滤纯化扩增子。由于这种方法需要进行几次操作，因此研究了替代的纯化程序。这些方法包括使用1）Qiagen QlAquick PCR纯化柱，2）Concert Rapid PCR纯化柱，以及3）ExoSAP-IT试剂。在评估纯化扩增子的产量、序列图谱质量和检测的简便性时，选择使用ExoSAP-IT试剂进行扩增后纯化来取代过滤方法。
