Rapid Screening for Five Major Cystic Fibrosis Mutations by Melting Peak Analysis n
Table 2. Melting temperatures of different alleles of the CFTR gene
Locus
Allele
Pairing
Tm (observed)
R347P
WT
C- G match
62 D C
Mutation
C- C mismatch
SIDC
.o.FS08
WT
TTT match
66 D C
Mutation
-- - Deletion
S7 D C
GS42X
WT
C-G match
63.5 D C
Mutation
C- T mismatch
S7 D C
GS51D
WT
C-G match
65 D C
Mutation
C-A mismatch
62 D C
R553X
WT
G-C match
65 D C
Mutation
G-T mismatch
59 D C
WT, wild-type sequence.
Table 3. Genotypes of the DNA samples used for CF mutation analysis
Sample
Mutation
R347P
L1F508
G542X
G55ID
R553X
I
2
HE
HE
WT
WT
WT
3
WT
HE
HE
WT
WT
4
WT
WT
WT
HE
WT
5
WT
WT
WT
WT
HE
WT, wild-type sequence; HE, heterozygous mutation.
dye, e.g., for L1FS08 and GS42X detection, show very similar T mS' Therefore it is
necessary to run the test in two separate capillaries. It may be possible to redesign
the probes in order to obtain distinguishable T mS for the probes labelled with the
same dye.
References
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