Modified salting-out method for DNA isolation from newborn cord blood nucleated cells.
The present work describes modification of a widely used salting-out procedure to rapidly extract DNA suitable for PCR, using the ARMS method to amplify a target sequence in the beta-globin gene. The salting-out DNA extraction procedure did not completely remove or decrease the presence of inhibitors to PCR in a considerable number of cord blood samples. By introducing a simple phenol/chloroform step, before ethanol precipitation of the nucleic acid, to certain samples, we were able to eliminate or substantially reduce the presence of inhibitors to PCR without having to re-extract the samples.
Noguera
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Cátedra y Servicio de Hematología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Hospital Provincial del Centenario, Rosario, Santa Fe, Argentina. nnofuwe@fbioyf.unr.edu.ar
Tallano
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Bragós
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Milani
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D001773:Blood Cells / Q000737:chemistry*; D011232:Chemical Precipitation; D002725:Chloroform; D004247:DNA / Q000097:blood*; D004587:Electrophoresis, Agar Gel; D000431:Ethanol; D004996:Ethidium; D005312:Fetal Blood / Q000166:cytology*; D005914:Globins / Q000235:genetics*; D006801:Humans; D007202:Indicators and Reagents; D007231:Infant, Newborn; D019800:Phenol; D016133:Polymerase Chain Reaction; D013053:Spectrophotometry
D007202:Indicators and Reagents; D019800:Phenol; D000431:Ethanol; D002725:Chloroform; D005914:Globins; D004247:DNA; D004996:Ethidium