Microgel electrophoresis: sensitivity, mechanisms, and DNA electrostretching.
Based on the treatment of microgels to remove proteins, we speculate that proteins may be bound to DNA in the microgels even after electrophoresis. We speculate that some DNA single-strand breaks may be a reflection of these protein-DNA complexes. We suggest methods to limit such artifacts, and present data demonstrating a lymphocyte DNA double-strand break sensitivity of 12.5 rads and day-to-day reproducibility of microgel electrophoresis using these principles. Extending these principles, we describe DNA behavior during alkaline and neutral microgel electrophoresis based on observations of the stained DNA and its migration patterns. During microgel electrophoresis, individual DNA molecules behave as if anchored at one end while the other end is free to migrate in response to the electric field. We capitalize on this behavior by developing a neutral microgel method to stretch chromosomes.
Singh
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NP
Bioelectromagnetics Research Laboratory, University of Washington, Seattle 98195-7962, USA. narendra@u.washington.edu
Stephens
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RE
D004249:DNA Damage; D004260:DNA Repair; D003851:Deoxyribonucleases / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D005782:Gels / Q000032:analysis; D006801:Humans; D008214:Lymphocytes; D011485:Protein Binding; D015203:Reproducibility of Results; D012260:Ribonucleases / Q000494:pharmacology; D012680:Sensitivity and Specificity
D005782:Gels; D003851:Deoxyribonucleases; D012260:Ribonucleases