Limitations of Melting Curve Analysis Using SYBR Green I - Fragment Differentiation and Mutation Detection .
Table 1. Continued
CFTR Accession No.: M55 \07, M551 15, M55116, M55127, M55129
CFTRexon
length
GC(%)
Tm (0e)
Primers
GAGCCTTCAGAGGGTAAAAT
20
45
58.4
GGGTTCATATGCATAATCAAA
21
33
55.3
Product 3
10
200
37
80.1
Primers
GTGAAIATCTGTTCCTCCTC
20
45
56.1
TATGTCTGACAATTCCAGGC
20
45
58.8
Product 4
2
90
42
80.2
Primers
ATGGAGACCAAATCAAGTGAA
21
38
58.6
TCAGCAGAATCAACAGAAGG
20
45
59.0
Product 5
2
135
42
80.9
Primers
TTCAAATGGTGGCAGGTAGT
20
45
60.7
CCACAAGGACAAAGTCAAGC
20
50
60.8
Product 6
22
200
42
82
Primers
TCTTCTCTAACTGCAGGTTG
20
45
58.3
CTGAAACTCACACTGGATCC
20
50
59.3
Product 7
22
200
49
86.3
Primers
TTATGCCTGGCACCATTAAA
20
40
59.0
CGCTTCTGTATCTATATTCATC
22
36
55.1
Product 8
]0
69
35
76
Primers
TTATGCCTGGCACCATTAAA
20
40
59.0
TTGATGACGCTTCTGTATCT
20
40
57.8
Product 9
10
76
34
77.8
Primers
GGATCAGGGAAGAGIACTT
19
47
56.6
GATTTCTCCTTCAGTGTTCA
20
40
56.4
Product 10
20
63
40
Primers
GGATCAGGGAAGAGIACTT
19
47
56.6
CGATCTGGATTTCTCCTTCA
20
45
58.2
Product 11
20
70
41
Primers
CTCCAAGTTTGCAGAGAAAG
20
45
58.1
CTCCACTCAGTGTGATTCC
19
53
58.7
Product 12
II
62
44
79.02
Melting temperatures (T m) of the oligonucleotides were calculated using the nearest-neighbour method (7). Melting temperatures of the products are mean experimental values.
Table 1. Continued
CFTR Accession No.: M55 \07, M551 15, M55116, M55127, M55129
CFTRexon
length
GC(%)
Tm (0e)
Primers
GAGCCTTCAGAGGGTAAAAT
20
45
58.4
GGGTTCATATGCATAATCAAA
21
33
55.3
Product 3
10
200
37
80.1
Primers
GTGAAIATCTGTTCCTCCTC
20
45
56.1
TATGTCTGACAATTCCAGGC
20
45
58.8
Product 4
2
90
42
80.2
Primers
ATGGAGACCAAATCAAGTGAA
21
38
58.6
TCAGCAGAATCAACAGAAGG
20
45
59.0
Product 5
2
135
42
80.9
Primers
TTCAAATGGTGGCAGGTAGT
20
45
60.7
CCACAAGGACAAAGTCAAGC
20
50
60.8
Product 6
22
200
42
82
Primers
TCTTCTCTAACTGCAGGTTG
20
45
58.3
CTGAAACTCACACTGGATCC
20
50
59.3
Product 7
22
200
49
86.3
Primers
TTATGCCTGGCACCATTAAA
20
40
59.0
CGCTTCTGTATCTATATTCATC
22
36
55.1
Product 8
]0
69
35
76
Primers
TTATGCCTGGCACCATTAAA
20
40
59.0
TTGATGACGCTTCTGTATCT
20
40
57.8
Product 9
10
76
34
77.8
Primers
GGATCAGGGAAGAGIACTT
19
47
56.6
GATTTCTCCTTCAGTGTTCA
20
40
56.4
Product 10
20
63
40
Primers
GGATCAGGGAAGAGIACTT
19
47
56.6
CGATCTGGATTTCTCCTTCA
20
45
58.2
Product 11
20
70
41
Primers
CTCCAAGTTTGCAGAGAAAG
20
45
58.1
CTCCACTCAGTGTGATTCC
19
53
58.7
Product 12
II
62
44
79.02
Melting temperatures (T m) of the oligonucleotides were calculated using the nearest-neighbour method (7). Melting temperatures of the products are mean experimental values.