Inhibition of DNA synthesis by K+-stabilised G-quadruplex promotes allelic preferential amplification.
PCR preferential amplification consists of the inefficient amplification of one allele in a heterozygous sample. Here, we report the isolation of a GC-rich human minisatellite, MsH43, that undergoes allelic preferential amplification during PCR. This effect requires the existence of a (TGGGGC)(4) motif that is able to form a G-quadruplex in the presence of K(+). This structure interferes with the DNA synthesis of the alleles harbouring this motif during PCR The present results are the first demonstration that the formation of G-quadruplex can be one of the mechanisms involved in some kinds of preferential amplification.
Departamento de Bioquímica y Biología Molecular, Facultad de Biología, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Galicia, Spain.
Francisco
F
Boán
Miguel G
MG
Blanco
Paula
P
Barros
Ana Isabel
AI
González
Jaime
J
Gómez-Márquez
D001483:Base Sequence; D019175:DNA Methylation; D004261:DNA Replication / Q000502:physiology*; D005260:Female; D005333:Fetus; D005784:Gene Amplification; D006801:Humans; D008099:Liver / Q000196:embryology / Q000502:physiology; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods; D011188:Potassium / Q000494:pharmacology*
D011188:Potassium