[47] The analysis of nucleic acids in gels using glyoxal and acridine orange

摘要: Publisher Summary This chapter discusses the analysis of nucleic acids in gels using glyoxal and acridine orange. To determine molecular weights gel electrophoresis techniques, molecules must have equivalent conformations. is achieved by removing native secondary tertiary structure with a variety chemical denaturants, thus reducing electrophoretic mobility to simple function polynucleotide weight, composition, voltage gradient applied. The describes method for denaturing RNA or DNA glyoxal, followed through either polyacrylamide- agarose-containing slab low ionic strength buffer. Using this reliable weight estimates varying sizes G + C contents were obtained; glyoxalated shown lie on same log versus curve, very similar curves. use metachromatic stain, orange, visualization gels. When intercalated between stacked bases double-helical acids, orange manifests green fluorescence. Its metachromasy visually confirms denaturation makes it powerful reagent rapid determination gross acid after electrophoresis.