SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells.
نویسندگان
چکیده
Pulsed field gel electrophoresis (PFGE) is widely used to measure DNA double strand breaks (dsb). The DNA of cultured cells can be prelabelled with radioactivity, which helps greatly in detection and quantitation of DNA dsb. However, this approach cannot be used with non-cycling cells from biopsy material. We describe a method which uses SYBR Green I to stain DNA in dried agarose gels. DNA is detected and analysed using readily available camera equipment and image analysis software. This method is as sensitive as [3H]thymidine prelabelling of cells and allows DNA dsb to be measured simply and economically in non-cycling cells.
منابع مشابه
Simulation of strand breaks induced in DNA molecule by radiation of proton and Secondary particles using Geant4 code
Radiotherapy using various beams is one of the methods for treating cancer, Hadrons used to treat cancers that are near critical organs. The most important part of the cell that is damage by ionizing radiation is DNA. In this study, damages induced in the genetic material of living cells (DNA) defined by the atomic model from the protein data bank (PDB) have been studied by radiati...
متن کاملModeling the distribution of deposited energy by alpha particles from Radon 223 decay and its effect on DNA
The ionizing radiations, through physical and chemical processes, lead to simple and complex single- and double- strand breaks, as well as base lesions to the DNA. In this study, taking into account all the physical and chemical processes involved in the interaction of ionizing radiation with matter, the initial damage induced to DNA was evaluated for 5.7 MeV alpha-rays from Radon 223 isotope....
متن کاملMethyl methanesulfonate (MMS) produces heat-labile DNA damage but no detectable in vivo DNA double-strand breaks
Homologous recombination (HR) deficient cells are sensitive to methyl methanesulfonate (MMS). HR is usually involved in the repair of DNA double-strand breaks (DSBs) in Saccharomyces cerevisiae implying that MMS somehow induces DSBs in vivo. Indeed there is evidence, based on pulsed-field gel electrophoresis (PFGE), that MMS causes DNA fragmentation. However, the mechanism through which MMS ind...
متن کاملEvaluating Gamma-H2AX Expression as a Biomarker of DNA Damage after X-ray in Angiography Patients
Objective: Coronary heart disease (CHD) is one of the most common diseases. Coronary angiography (CAG) is an important apparatus used to diagnose and treat this disease. Since angiography is performed through exposure to ionizing radiation, it can cause harmful effects induced by double-stranded breaks in DNA which is potentially life-threatening damage. The aim of the present study is to inves...
متن کاملInduction of Apoptosis on K562 Cell Line and Double Strand Breaks on Colon Cancer Cell Line Expressing High Affinity Receptor for Granulocyte Macrophage-Colony Stimulating factor (GM-CSF)
Background: Immunotoxins are comprised of both the cell targeting and the cell killing moieties. We previously established a new immunotoxin, i.e. Shiga toxin granulocyte macrophage-colony stimulating factor (StxA1-GM-CSF), comprises of catalytic domain of Stx, as a killing moiety and GM-CSF, as a cell targeting moiety. In this study, the ability of the immunotoxin to induce apoptosis and dou...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Nucleic acids research
دوره 25 14 شماره
صفحات -
تاریخ انتشار 1997