Liu Yuxuan, He Huayu, Yi Shaohua, Hu Qingqing, Zhang Wenqiong, Huang Daixin
Department of Forensic Medicine, Tongji Medical College, Huazhong University of Science and Technology, No. 13, Hangkong Road, Wuhan, 430030, China.
Criminal Investigation Team, Sui County Public Security Bureau, Suizhou, 441300, China.
Int J Legal Med. 2018 May;132(3):675-681. doi: 10.1007/s00414-017-1666-7. Epub 2017 Aug 30.
Formalin fixation is considered an important process for preservation of human tissue samples for long periods. However, this process not only results in cross-linking complicating isolation of nucleic acid but also introduces polymerase "blocks" during polymerase chain reaction (PCR). At present, many protocols have already been developed aiming at extracting high amounts of amplifiable DNA from formalin-fixed tissues (FFTs). However, there are few methods for repairing formalin-damaged DNA. In this study, we compared the effectiveness of several post-extraction enzymatic repair techniques, including Taq DNA polymerase, DNA polymerase I and T4 DNA ligase, the PreCR™ Repair Mix and Restorase® DNA Polymerase, in restoring STR profiles from formalin-damaged DNA. Our results indicated that formalin-damaged DNA may be repaired partly with Taq DNA polymerase and the Restorase® DNA Polymerase, and lost alleles may be restored and STR peak heights may increase upon repair with them. Moreover, the repair ability of the protocol 2 with Taq DNA polymerase surpasses the Restorase® DNA Polymerase.
福尔马林固定被认为是长期保存人体组织样本的重要过程。然而,这一过程不仅会导致交联,使核酸分离变得复杂,还会在聚合酶链反应(PCR)过程中引入聚合酶“阻断”。目前,已经开发出许多方案,旨在从福尔马林固定组织(FFT)中提取大量可扩增的DNA。然而,修复福尔马林损伤DNA的方法却很少。在本研究中,我们比较了几种提取后酶修复技术的有效性,包括Taq DNA聚合酶、DNA聚合酶I和T4 DNA连接酶、PreCR™修复混合物和Restorase® DNA聚合酶,用于从福尔马林损伤的DNA中恢复STR图谱。我们的结果表明,福尔马林损伤的DNA可以用Taq DNA聚合酶和Restorase® DNA聚合酶部分修复,修复后丢失的等位基因可能会恢复,STR峰高可能会增加。此外,使用Taq DNA聚合酶的方案2的修复能力超过Restorase® DNA聚合酶。