School of Molecular Biosciences, Washington State University, Pullman, WA 99164, USA.
Center for Reproductive Biology, Washington State University, Pullman, WA 99164, USA.
STAR Protoc. 2021 Dec 22;3(1):101059. doi: 10.1016/j.xpro.2021.101059. eCollection 2022 Mar 18.
Exposure to ultraviolet (UV) light induces DNA damage, predominantly cyclobutane pyrimidine dimers (CPD) and 6,4-photoproducts (6,4-PP), as well as rare, atypical photoproducts at thymidine-adenine (TA) sequences. We have recently shown 'TA' photoproducts are induced in UV-irradiated oligonucleotides and across the budding yeast genome. Here, we describe a protocol for mapping atypical 'TA' photoproducts and This protocol overcomes the technical challenges involved in accurately mapping such rare photoproducts by using ultraviolet damage endonuclease (UVDE) enzymes. For complete details on the use and execution of this protocol, please refer to Laughery et al. (2020).
暴露于紫外 (UV) 光会诱导 DNA 损伤，主要是环丁烷嘧啶二聚体 (CPD) 和 6,4-光产物 (6,4-PP)，以及在胸腺嘧啶-腺嘌呤 (TA) 序列处罕见的、非典型的光产物。我们最近表明，在紫外线照射的寡核苷酸中和芽殖酵母基因组中都会诱导产生“TA”光产物。在这里，我们描述了一种用于绘制非典型“TA”光产物图谱的方案。该方案通过使用紫外线损伤内切酶 (UVDE) 酶克服了准确绘制此类稀有光产物所涉及的技术挑战。有关此方案的使用和执行的完整详细信息，请参阅 Laughery 等人 (2020)。
