Application of a novel fluorescence probe in the determination of nucleic acids.
A novel fluorimetric method has been developed for rapid determination of DNA and RNA with hypocrellin A (HA) as a fluorescence probe, based on the fluorescence enhancement of HA in the presence of DNA or RNA. Maximum fluorescence is produced in the pH range 3.4-4.0, with maximum excitation and emission wavelengths at 470 and 600 nm, respectively. Under optimal conditions, the calibration graphs are linear over the range 0-200.0 ng cm-3 for calf thymus DNA and 13.0-200.0 ng cm-3 for yeast RNA, respectively. The corresponding detection limits are 5.0 ng cm-3 for calf thymus DNA and 13.0 ng cm-3 for yeast RNA. The relative standard deviation of six replicate measurements is 4.5% for 100 ng cm-3 calf thymus DNA. DNA could be determined in the presence of 20% m/m yeast RNA. The mechanism for the binding of HA to DNA is also studied.
Zhu
Q Z
QZ
Department of Chemistry, Xiamen University, China.
Li
F
F
Guo
X Q
XQ
Xu
J G
JG
Li
W Y
WY
D004247:DNA / Q000032:analysis*; D005456:Fluorescent Dyes; D006801:Humans; D010569:Perylene / Q000031:analogs & derivatives; D019800:Phenol; D017319:Photosensitizing Agents; D011809:Quinones; D012313:RNA / Q000032:analysis*; D013050:Spectrometry, Fluorescence / Q000379:methods
D005456:Fluorescent Dyes; D017319:Photosensitizing Agents; D011809:Quinones; D019800:Phenol; D010569:Perylene; D012313:RNA; D004247:DNA; C048388:hypocrellin A