Inclusion of synthetic DNA templates of similar length and base composition to PCR-amplified products in restriction enzyme digestions: an efficient aid in characterization of point mutations.
Because of a subtle anomaly we encountered upon an analytical gel while characterizing a point mutation in an exon of a patient, we decided to perform expensive and time-consuming procedures to characterize the anomaly. Although initial and subsequent Southern blots and PCR analyses of this patient's mutation suggested that his mutation lay directly within a TaqI recognition site, further characterization revealed that the mutation actually lay in a base immediately outside the recognition site. Had we included an appropriate double-stranded DNA control in the restriction enzyme digestion of this patient's PCR-amplified exon, we could have arrived at the correct conclusion as to the location of the mutation without incurring high costs and time loss. This brief report depicts the use of DNA controls of appropriate length and base composition as a means of avoiding erroneous conclusions and expense in routine mutational analyses in the clinical setting.
Klein
D
D
Division of Endocrinology, Harbor-UCLA Medical Center, Torrance 90502.
Tabor
D E
DE
Sokol
R Z
RZ
Cederbaum
S D
SD
D000483:Alleles; D001482:Base Composition; D004247:DNA; D004252:DNA Mutational Analysis / Q000379:methods*; D004262:DNA Restriction Enzymes; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D013698:Templates, Genetic
D004247:DNA; D004262:DNA Restriction Enzymes