The protein effect on determination of DNA with Hoechst 33258.
The present study was designed to afford a critical review of the effect of proteins on the Hoechst 33258 method for determination of DNA of crude homogenates. A considerable effect of proteins on the fluorescence was observed when the concentration exceeded 100 micrograms BSA equivalent protein. Below that value, practically no effect of proteins was noted. We used proteinase K to remove the proteins, but dilution of homogenates could be used as well. Moreover, we found that the concentration of the fluorochrome should be between 1 microgram and 2 micrograms when microgram levels DNA are to be determined.
Moe
D
D
Department of Oral Function, Dental School, Faculty of Health Sciences, University of Copenhagen, Denmark.
Garbarsch
C
C
Kirkeby
S
S
D000818:Animals; D016477:Artifacts; D006690:Bisbenzimidazole; D004247:DNA / Q000032:analysis*; D003851:Deoxyribonucleases; D019286:Endopeptidase K; D005260:Female; D005453:Fluorescence; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D012697:Serine Endopeptidases; D012710:Serum Albumin, Bovine
D012710:Serum Albumin, Bovine; D004247:DNA; D003851:Deoxyribonucleases; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D006690:Bisbenzimidazole