N,N'-bis[3,3'-(dimethylamino)propylamine]-3,4,9, 10-perylenetetracarboxylic diimide, a dicationic perylene dye for rapid precipitation and quantitation of trace amounts of DNA.
A novel dicationic dye with a polycyclic aromatic perylene core and flexible cationic side chains- N,N'-bis[3, 3'-(dimethylamino)propylamine]-3,4,9,10-perylenetetracarboxylic diimide-termed "DAPER," was synthesized and characterized. The dye appears to exist in a highly stacked form in aqueous solution. DAPER precipitates extremely low concentrations of DNA very rapidly, efficiently, and with a stoichiometry of one tightly bound dye per DNA base pair, corresponding to a neutral complex. Precipitation may occur due to side-by-side association between the polyanionic DNA helix and polycationic dye stacks. DNA precipitation by DAPER is less sensitive to DNA concentration and length, and prevailing salt concentrations, than precipitation with ethanol or propanol. DAPER can be quantitatively extracted from DNA into a standard phenol:chloroform mixture under slightly alkaline conditions. The recovered DNA is suitable for treatment with enzymes typically used in DNA sequencing procedures. The amount of DNA precipitated is accurately determined by visible absorption or fluorescence spectroscopic analyses of the phenol:chloroform extracts. Several samples of DNA can be precipitated, recovered, and quantitated in about 1 h using standard microscale procedures and equipment. The unique qualities of DAPER provide the basis for a very sensitive, rapid, and versatile method for simultaneous precipitation and quantitation of microgram and submicrogram amounts of DNA.
Liu
Z R
ZR
Department of Chemistry and Institute of Molecular Biophysics, Florida State University, Tallahassee, 32306-3006, USA.
Rill
R L
RL
D000433:1-Propanol / Q000737:chemistry; D011232:Chemical Precipitation; D004396:Coloring Agents; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D007094:Imides / Q000737:chemistry*; D010569:Perylene / Q000031:analogs & derivatives* / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
D004396:Coloring Agents; D007094:Imides; C099239:N,N'-bis(3,3'-(dimethylamino)propylamine)-3,4,9,10-perylenetetracarboxylic diimide; D010569:Perylene; D004247:DNA; D000433:1-Propanol