PNA-mediated purification of PCR amplifiable human genomic DNA from whole blood.
A simple and rapid procedure whereby human genomic DNA can be purified in a PCR amplifiable form from whole blood is described. In a first step, human genomic DNA is hybridized in solution to a biotinylated peptide nucleic acid (PNA), which forms a high-affinity triplex with A7 sequence motifs in the target DNA. The complex is then captured onto paramagnetic streptavidin-coated particles, which are subsequently transferred directly into the PCR. The purification method effectively removes inhibitors of the PCR from as much as 500 microL of whole blood.
Seeger
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PNA Diagnostics A/S Copenhagen, Denmark.
Batz
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D001710:Biotin; D002878:Chromosomes, Human, Pair 1; D002897:Chromosomes, Human, Pair 7; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004247:DNA / Q000097:blood / Q000302:isolation & purification*; D005164:Factor IX / Q000235:genetics; D005260:Female; D006639:Histidine; D006801:Humans; D008297:Male; D009693:Nucleic Acid Hybridization; D009696:Nucleic Acids; D010455:Peptides; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D014960:X Chromosome
C505032:CFTR protein, human; D009696:Nucleic Acids; D010455:Peptides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D006639:Histidine; D001710:Biotin; D005164:Factor IX; D004247:DNA