Amplification Primers for DGGE-Based Mutation Analysis of the Fas Gene
| Exon/ Primer . | Sequence (5′ → 3′)* . | Product Length (bp) . | Tm of Domain (°C)-151 . |
|---|---|---|---|
| FAS-734GC | [GC]-TCCCTTTTCAGAGCCCTATGG | 178 | 75.6 |
| FAS-623 | GACTTGCGGGGCATTTGAC | ||
| 1-F-152 | cgccg[GC]-TCAGTACGGAGTTGGGGAAGC | 182 | 76.9 |
| 1-R | GCCTATCCCCGGGACTAAGAC | ||
| 2-F | [GC]-ATCAATAAAATTCTCTTCATGC | 190 | 68.9 |
| 2-R | TGACTTTCACTGTAATCTCTGG | ||
| 3-F | AAACACTTGCTCCTTTTTTCC | 254 | 71.7 |
| 3-R | [GC]-TGAAATTCCAAGATTGGCC | ||
| 4-F | gcccgTCCAAACTGATTTTCTAGGC | 207 | 69.5 |
| 4-R | [GC]-TCTAGTGTTTTAATCAGAGAAAGAC | ||
| 5-F | [GC]-CCAGGCTTTTGAATTTCTCC | 182 | 69.1 |
| 5-R | ccgcgccgGGGAAAGGAGGATATAACCG | ||
| 6-F | ATAATATGCCAATGTTCCAACC | 175 | 69.4 |
| 6-R | [GC]-CCCCAAGTTATTTCAATCTGC | ||
| 7-F | CATGCATTCTACAAGGCTGAG | 255 | 67.2 |
| 7-R | [GC]-AGGAAGTAACAAAAAGCCAAATC | ||
| 8-F | TCTCTGCTTCCATTTTTTGC | 159 | 65.4 |
| 8-R | [GC]-TTTACTCTGAAATTGGCCTATTAC | ||
| 9I-F | [GC]-TATTTTCTATTTTTCAGATGTTGAC | 275 | 67.9 |
| 9I-R | TCATACGCTTCTTTCTTTCC | ||
| 9II-F | GTTCAACTGCTTCGTAATTG | 251 | 67.6 |
| 9II-R | [GC]-AGAACTGAATTTGTTGTTTTTC |
| Exon/ Primer . | Sequence (5′ → 3′)* . | Product Length (bp) . | Tm of Domain (°C)-151 . |
|---|---|---|---|
| FAS-734GC | [GC]-TCCCTTTTCAGAGCCCTATGG | 178 | 75.6 |
| FAS-623 | GACTTGCGGGGCATTTGAC | ||
| 1-F-152 | cgccg[GC]-TCAGTACGGAGTTGGGGAAGC | 182 | 76.9 |
| 1-R | GCCTATCCCCGGGACTAAGAC | ||
| 2-F | [GC]-ATCAATAAAATTCTCTTCATGC | 190 | 68.9 |
| 2-R | TGACTTTCACTGTAATCTCTGG | ||
| 3-F | AAACACTTGCTCCTTTTTTCC | 254 | 71.7 |
| 3-R | [GC]-TGAAATTCCAAGATTGGCC | ||
| 4-F | gcccgTCCAAACTGATTTTCTAGGC | 207 | 69.5 |
| 4-R | [GC]-TCTAGTGTTTTAATCAGAGAAAGAC | ||
| 5-F | [GC]-CCAGGCTTTTGAATTTCTCC | 182 | 69.1 |
| 5-R | ccgcgccgGGGAAAGGAGGATATAACCG | ||
| 6-F | ATAATATGCCAATGTTCCAACC | 175 | 69.4 |
| 6-R | [GC]-CCCCAAGTTATTTCAATCTGC | ||
| 7-F | CATGCATTCTACAAGGCTGAG | 255 | 67.2 |
| 7-R | [GC]-AGGAAGTAACAAAAAGCCAAATC | ||
| 8-F | TCTCTGCTTCCATTTTTTGC | 159 | 65.4 |
| 8-R | [GC]-TTTACTCTGAAATTGGCCTATTAC | ||
| 9I-F | [GC]-TATTTTCTATTTTTCAGATGTTGAC | 275 | 67.9 |
| 9I-R | TCATACGCTTCTTTCTTTCC | ||
| 9II-F | GTTCAACTGCTTCGTAATTG | 251 | 67.6 |
| 9II-R | [GC]-AGAACTGAATTTGTTGTTTTTC |
*Lowercase characters represent nucleotides incorporated into the primer to modulate the melting profile of the amplification product.
Melting temperature of the lower melting domain of the GC-clamped PCR product, as determined by MELT87.16
F, forward primer; R, reverse primer; [GC], CGCCCGCCGCGCCCCGCGCCCGTCCCGCCGCCCCCGCCCG.