- 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
PMID34401774
ABSTRACT
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).
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Dulbecco′s Modified Eagle′s Medium - low glucose, With 1000 mg/L glucose, L-glutamine and sodium bicarbonate, without phenol red, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Dulbecco′s Phosphate Buffered Saline, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, fatty acid free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
Penicillin-Streptomycin, 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
L-Glutamine solution, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture
Millipore
Triton™ X-100 Detergent, Molecular Biology Grade, Non-ionic detergent and emulsifier. Has an optimal pH range of 6.0-8.0.