Detection of low dose radiation induced DNA damage using temperature differential fluorescence assay.
A rapid and sensitive fluorescence assay for radiation-induced DNA damage is reported. Changes in temperature-induced strand separation in both calf thymus DNA and plasmid DNA (puc 19 plasmid from Escherichia coli) were measured after exposure to low doses of radiation. Exposures of between 0.004 and 1 Gy were measured with doses as low as 0.008 Gy yielding significant responses. The double-strand, sensitive dye PicoGreen was used as an indicator of DNA denaturation. Calibration plots indicate that fluorescence changes corresponding to amounts as low as 1 ng of double stranded DNA (10(6) copies for plasmid puc 19) are detected by this method.
Rogers
K R
KR
Apostol
A
A
Madsen
S J
SJ
Spencer
C W
CW
D000818:Animals; D002417:Cattle; D004247:DNA / Q000032:analysis / Q000528:radiation effects*; D004249:DNA Damage; D004269:DNA, Bacterial / Q000032:analysis / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004926:Escherichia coli / Q000235:genetics; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D005470:Fluorometry / Q000379:methods*; D066298:In Vitro Techniques; D009930:Organic Chemicals; D010957:Plasmids / Q000235:genetics; D011833:Radiation Injuries, Experimental / Q000175:diagnosis*; D011839:Radiation, Ionizing; D012680:Sensitivity and Specificity; D013696:Temperature
D004269:DNA, Bacterial; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C099476:PicoGreen; D004247:DNA