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  • 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
要旨

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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製品内容

Sigma-Aldrich
ダルベッコリン酸緩衝生理食塩水, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
ウシ血清アルブミン ウシ血清由来, heat shock fraction, protease free, fatty acid free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
L-グルタミン 溶液, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
ペニシリン-ストレプトマイシン, 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-ブロモ-2′-デオキシウリジン, BioUltra, ≥99%
Millipore
Triton X-100, Non-ionic detergent and emulsifier. Has an optimal pH range of 6.0-8.0.
Sigma-Aldrich
ダルベッコ改変イーグル培地 - 低グルコース, With 1000 mg/L glucose, L-glutamine and sodium bicarbonate, without phenol red, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
ビスベンズイミド H 33342 三塩酸塩, ≥98% (HPLC and TLC)