Comparison of different methods for repairing damaged DNA from buffered and unbuffered formalin-fixed tissues
Comparison of different methods for repairing damaged DNA from buffered and unbuffered formalin-fixed tissues
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缓冲和未缓冲福尔马林固定组织中修复受损 DNA 的不同方法的比较
DOI：
10.1007/s00414-017-1666-7
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发表时间：
2018-05-01
影响因子：
2.1
通讯作者：
Huang, Daixin
中科院分区：
文献类型：
--
作者：
Liu, Yuxuan;He, Huayu;Huang, Daixin
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，该过程不仅导致使核酸分离复杂化的交联，而且在聚合酶链式反应（PCR）期间引入聚合酶“阻断”。目前，已经开发了许多方案，旨在从福尔马林固定的组织（FFT）中提取大量的可扩增DNA。然而，几乎没有修复福尔马林损伤的DNA的方法。在这项研究中，我们比较了几种提取后酶修复技术，包括Taq DNA聚合酶，DNA聚合酶I和T4 DNA连接酶，PreCR（TM）Repair Mix和RestoraseA（R）DNA聚合酶，在恢复福尔马林损伤DNA的STR图谱中的有效性。我们的结果表明，Taq DNA聚合酶和RestoraseA（R）DNA聚合酶可以部分修复福尔马林损伤的DNA，修复后丢失的等位基因可以恢复，STR峰高可以增加。此外，使用Taq DNA聚合酶的方案2的修复能力优于RestoraseA（R）DNA聚合酶。
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 (TM) Repair Mix and RestoraseA (R) 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 RestoraseA (R) 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 RestoraseA (R) DNA Polymerase.