Biology Reference
In-Depth Information
FIGURE 3.4. SSCP analysis of PCR prod-
ucts of exon 2 of the coding region of the
Cx26
gene of DNA samples from persons
with nonsyndromic hearing impairment
showing a mobility shift/doublet in an indi-
vidual with the 310de114 mutation in the
Cx26
gene in the sample in Lane 2.
in mobility from homoduplexes on polyacrylamide gel analysis (Nagamine
et al. 1989). This is thought to be due to sequence-dependent conforma-
tional changes in the double-stranded DNA duplex. Heteroduplex analysis
has a similar sensitivity to SSCP for DNA fragments less than 300 bp in size
(White et al. 1992).
8.2.4 Cleavage Mismatch
Cleavage mismatch involves adding the chemicals hydroxylamine and
osmium tetroxide, which react with free cytosine and thymine nucleotides,
respectively. The DNA template is denatured, then hybridized with a single-
stranded radiolabeled DNA probe. Any mismatched cytosine or thymine
nucleotides will be exposed and susceptible of reaction with the hydroxy-
