DNA elution and amplification by polymerase chain reaction from dried blood spots.
A quick, sensitive and easily automatizable method for PCR amplification of genomic DNA eluted from dried blood spots is described. DNA is eluted from a 3-mm spot routinely used for neonatal screening of inherited diseases either by boiling or by sonication. A preliminary and brief spot-autoclaving step is mandatory to ensure optimal and reproducible PCR amplifications. Only 1% of the eluted DNA is required for PCR analysis allowing the execution of multiple genetic tests on the same blood spot. The method has been successfully applied to the detection of a known phenylketonuria-causing mutation and will facilitate the analysis of the genetic repository provided by Guthrie's cards stored in neonatal screening laboratories.
Carducci
C
C
Dept. of Experimental Medicine, University of Rome La Sapienza, Italy.
Ellul
L
L
Antonozzi
I
I
Pontecorvi
A
A
D001483:Base Sequence; D004247:DNA / Q000097:blood*; D015342:DNA Probes; D005260:Female; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*
D015342:DNA Probes; D004247:DNA