Table 1 Primers and pyrosequencing assay information. Primers sequences used for PCR, Sanger sequencing, pyrosequencing and AFLP technique
From: Variation of mutant allele frequency in NRAS Q61 mutated melanomas
NRAS Pyrosequencing ASSAY 1 | |
PCR 124 nt | |
Primer F-Biotine | ACACCCCCAGGATTCTTACAGA |
Primer R | GCCTGTCCTCATGTATTGGTC |
Pyrosequencing primer | CATGGCACTGTACTCTTC |
Nucleotide injection order | GTTACGTCAGCTG |
NRAS Pyrosequencing ASSAY 2 | |
PCR 124 nt | |
Primer F-Biotine | ACACCCCCAGGATTCTTACAGA |
Primer R | GCCTGTCCTCATGTATTGGTC |
Pyrosequencing primer | CATGGCACTGTACTCTTC |
Nucleotide injection order | GCATACGTCAG |
NRAS Pyrosequencing ASSAY 3 | |
PCR 90 nt | |
Primer F-Biotine | ACAAGTGGTTATAGATGGTGA |
Primer R | ATGTATTGGTCTCTCATGGCA |
Pyrosequencing primer | CATGGCACTGTACTCTTC |
Nucleotide injection order | GCATACGTCAGCT |
Sanger Sequencing | |
Primer F | ACAAGTGGTTATAGATGGTGA |
Primer R | ATGTATTGGTCTCTCATGGCA |
AFLP primers | |
rs3219599 F | TTCAAGGCTGCAGTGAGCTA |
rs3219599 R | AGTGGAAGCTAGACACACATTAAGA |
s3219653 F | CCAGAGAGACAGAACTGAACAAA |
s3219653 R | CAAATTTTGGACCTGCCATG |
rs3219587 F | GGGCAAATGGAGGAAAGAGA |
rs3219587 R | TAAAAATACCCCCACCCCACT |
rs3220698 F | TTAAAAAACGTACTGCCACATTCA |
rs3220698 R | GGCAGAAACCAGGAAATGTAGTA |
rs3220987 F | GGCTTTTAGCTATGATTTGAGA |
rs3220987 R | GACTCAGGAAATAAACAAGGC |
rs3220389 F | CGCTGCTCACTCCTCCTCTGA |
rs3220389 R | AGTGCTGCTCTCAGTGAACTC |
rs3219612 F | AGCACACAATATACTCTCTCAGA |
rs3219612 R | ACCTGGGCAAAAGAGTAAGACC |
rs3219703 F | AACGAAGGTGTACTGGGACTGGT |
rs3219703 R | ACAGGGATGTGAGGGATTTTTTC |