DNA samples from different individuals of a family were pooled, amplified, and analyzed by CDCE, or the DNA from each individual was amplified and analyzed separately on the CDCE.
The optimal temperature for the resolution of fragment 13b was obtained by performing CDCE runs at different temperatures.
The detection limit of CDCE under the conditions used for the resolution of fragment 13b was next determined using DNA samples amplified from a homozygous (1987G) individual and a 1987A [right arrow] G heterozygote.
To evaluate the reproducibility of CDCE, we analyzed a sample containing a mixture of three mutants (50% 1853G [right arrow] T heterozygotes,16.
Analysis of the CDCE profiles of fragment 13a in the 31 families showed that the profiles for fragment 13a were similar to the standard Scnn1a sequence.
Regarding fragmented 13b, CDCE analysis of the 31 families showed that they could be categorized according to the observed peaks (CDCE profiles) into four groups.
The other 13 families clearly differed in their CDCE profiles from the standard sequence, indicating the presence of Scnn1a exon 13 variants.
Two families (families 17 and 19) had CDCE profiles containing three homoduplex peaks (peaks 1-3; Fig.
To interpret the various peaks observed in the CDCE profiles, we characterized the individuals of one family (family 17) from group 2 by CDCE and DNA sequence analysis.
The DNA from all members of families showing CDCE profiles consistent with the presence of ENaC variants were analyzed individually.
CDCE was used to screen for mutations in the COOH terminus of the [beta] subunit of ENaC (Scnn1b).
In this report, we show the application of CDCE for mutation detection in families participating in a study on the genetics of hypertension.