Table 1 of
Gamundi, Mol Vis 2005;
11:922-928.Table 1 of
Gamundi, Mol Vis 2005;
11:922-928.
Table 1. Primers, amplification, and DGGE conditions
GC-clamp corresponds to the sequence 5'-CGC CCG CCG CGC CCC GCG CCC GGC CCG CCG CCC CCG CCC G-3'. 100% denaturant is 7 M urea, 40% v/v formamide in TAE buffer. Gradient denaturation was permormed at 60 °C.
Fragment PCR annealing Percentage
size temperature DGGE gradient
Exon Primers (5'-3') (bp) (°C)/time(s) denaturant
---- --------------------------------- -------- ------------- -------------
1A F: gcgggggccgtgagcactca 383 60/60 55-95%
R: GC-clamp-caggaccaggaagcggcagt
1B F: GC-clamp-cgctacctgtcggcagaagag 415 58/30 55-100%
R: gctgtcacaggacttgaggcagta
1C F: GC-clamp-gacgccctcctcaccctcatc 426 58/30 55-90%
R: ccccacggcctcctccctgaa
2 F: GC-clamp-ggagaggcgtgaggggcttc 262 58/30 40-70%
R: ataggaggaagggatgtgtg
3 F: GC-clamp-cacccccatctcctgtc 236 65/60 40-80%
R: agtacctggcaggcagagtg
4 F: GC-clamp-gggaggggcagcgcagcaga 368 65/60 55-100%
R: ctgccgtggctgcccgtgta
5 F: ggggtggcagcgggcaggt 408 68/60 70-100%
R: GC-clamp-gggcctcctccacctccag
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