Analysis of DNA fragmentation using a dynamic size-sieving polymer solution in capillary electrophoresis.
Various natural and induced processes cause DNA fragmentation. Examples of these processes include apoptosis, enzymatic digestion, free radical production from ionizing radiation, photoscission by laser radiation and thermal degradation. Slab gel electrophoresis has been used most often to monitor such DNA damage. We have investigated with capillary electrophoresis the use of a new size-sieving polymer solution, TreviSol-CE (TS-CE), to monitor the DNA fragments produced from a variety of degradation processes. This polymer solution provides high run-to-run migration time and peak width reproducibilities and high separation efficiency of double-stranded DNA fragments in the 500 to 7000 base pair size range. Analysis of apoptotic DNA fragments suggested the presence of multiple nucleosomes within each cell type investigated. For irradiated DNA standards, peak-width-at-half-height and peak area were used to monitor the progress of DNA fragmentation. For both apoptotic DNA and irradiated DNA standards, fine structural features of fragmentation were revealed.
Siles
B A
BA
Analytical Chemistry Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.
Nackerdien
Z E
ZE
Collier
G B
GB
D017209:Apoptosis; D053938:DNA Fragmentation; D004273:DNA, Neoplasm / Q000032:analysis* / Q000187:drug effects / Q000235:genetics; D003851:Deoxyribonucleases / Q000494:pharmacology; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D005720:Gamma Rays; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000473:pathology; D010316:Particle Size; D011108:Polymers; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D011108:Polymers; D003851:Deoxyribonucleases