| Chapter title |
Fast Micromethod: Determination of DNA Integrity in Cell Suspensions and in Solid Tissues
|
|---|---|
| Chapter number | 13 |
| Book title |
Fast Detection of DNA Damage
|
| Published in |
Methods in molecular biology, July 2017
|
| DOI | 10.1007/978-1-4939-7187-9_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7185-5, 978-1-4939-7187-9
|
| Authors |
Nevenka Bihari |
| Abstract |
The Fast Micromethod is a rapid and convenient microplate procedure for the determination of DNA integrity in cell suspensions and in solid tissues. The procedure is based on the ability of fluorochromes to preferentially interact with double-stranded DNA in alkaline conditions. Rapid sample lysis is followed by denaturation at high pH during 15 min. Only 30 ng of DNA from cell suspensions or tissue homogenates per single well are required for the analyses. The whole analysis is performed within 3 h or less (for one 96-well microplate).The Fast Micromethod is broadly used in biology and medicine. Its applications range from environmental pollution tests in marine invertebrates to the analysis of biopsy samples in cancer patients to detect DNA alterations caused by irradiation or chemotherapy.The procedure presented here describes the Fast Micromethod applied for the determination of DNA integrity in cell suspensions (HeLa cells) and solid tissues (mussel gills). |
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