INCLUSION OF SYNTHETIC DNA TEMPLATES OF SIMILAR LENGTH AND BASE COMPOSITION TO PCR-AMPLIFIED PRODUCTS IN RESTRICTION ENZYME DIGESTIONS - ANEFFICIENT AID IN CHARACTERIZATION OF POINT MUTATIONS
D. Klein et al., INCLUSION OF SYNTHETIC DNA TEMPLATES OF SIMILAR LENGTH AND BASE COMPOSITION TO PCR-AMPLIFIED PRODUCTS IN RESTRICTION ENZYME DIGESTIONS - ANEFFICIENT AID IN CHARACTERIZATION OF POINT MUTATIONS, Somatic cell and molecular genetics, 20(1), 1994, pp. 61-65
Because of a subtle anomaly we encountered upon an analytical gel whil
e 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 analy
ses of this patient's mutation suggested that his mutation lay directl
y within a TaqI recognition site, further characterization revealed th
at the mutation actually lay in a base immediately outside the recogni
tion site. Had we included an appropriate double-stranded DNA control
in the restriction enzyme digestion of this patient's PCR-amplified ex
on, we could have arrived at the correct conclusion as to the location
of the mutation without incurring high costs and time loss. This brie
f report depicts the use of DNA controls of appropriate length and bas
e composition as a means of avoiding erroneous conclusions and expense
in routine mutational analyses in the clinical setting.