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Merck
  • A laser-mediated photo-manipulative toolbox for generation and real-time monitoring of DNA lesions.

A laser-mediated photo-manipulative toolbox for generation and real-time monitoring of DNA lesions.

STAR protocols (2021-08-18)
Bindhu K Madhavan, Zhe Han, Albert Sickmann, Rainer Pepperkok, Peter P Nawroth, Varun Kumar
摘要

With the advancement of laser-based microscopy tools, it is now possible to explore mechano-kinetic processes occurring inside the cell. Here, we describe the advanced protocol for studying the DNA repair kinetics in real time using the laser to induce the DNA damage. This protocol can be used for inducing, testing, and studying the repair mechanisms associated with DNA double-strand breaks, interstrand cross-link repair, and single-strand break repair. For complete details on the use and execution of this protocol, please refer to Kumar et al. (2017, 2020).

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Sigma-Aldrich
杜氏磷酸盐缓冲盐水, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
牛血清白蛋白 来源于牛血清, heat shock fraction, protease free, fatty acid free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
L -谷氨酰胺 溶液, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
青链霉素, with 10,000 units penicillin and 10 mg streptomycin per mL in 0.9% NaCl, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
5-溴-2′-脱氧尿苷, BioUltra, ≥99%
Millipore
TritonX-100洗涤剂,分子生物学级, Non-ionic detergent and emulsifier. Has an optimal pH range of 6.0-8.0.
Sigma-Aldrich
双苯并咪唑 H 33342 三盐酸盐, ≥98% (HPLC and TLC)
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杜氏改良 Eagle 培养基-低葡萄糖, With 1000 mg/L glucose, L-glutamine and sodium bicarbonate, without phenol red, liquid, sterile-filtered, suitable for cell culture