Table 2 Primer sequences and PCR amplification conditions for Sanger Sequencing (SS) validation
Gene | Chromosome Position | RefSeq | Coding DNA | Protein | PCR primers | Ta (°C) | Length amplicon (bp) |
|---|---|---|---|---|---|---|---|
CDKN2A | chr9:21974792 | NM_001195132 | c.35delC | p.(Ser12TrpfsTer14) | F: ACTTCAGGGGTGCCACATTC | 60 | 493 |
R: GCGCTACCTGATTCCAATTC | |||||||
TP53 | chr17:7579472 | NM_000546.5 | c.215C > G | p.Pro72Arg | F: TGAAGCTCCCAGAATGCCAG | 60 | 136 |
R: GCTGCCCTGGTAGGTTTTCT | |||||||
TP53 | chr17:7577543 | NM_000546.5 | c.738G > A | p.Met246Ile | F: TGGCTCTGACTGTACCACCA | 60 | 123 |
R: CAAGTGGCTCCTGACCTGG | |||||||
ERBB4 | chr2:212812278 | NM_005235 | c.298G > A | p.Glu100Ly | F: ACAGGCTACGTGTTAGTGGC | 60 | 104 |
R: GCCAAGGCATATCGATCCTCA | |||||||
ERBB4 | chr2:212578373 | NM_005235 | c.884A > T | p.His295Leu | F: TGTTTTGAGCTTGTTTGCTGA | 60 | 176 |
R: GGGCAAATGTCAGTGCAAGG | |||||||
ARID2 | chr12:46244997 | NM_152641 | c.3091C > T | p.Gln1031Ter | F: CGTCGTCCTCTACCCCTCAA | 60 | 201 |
R: CACCAGAGGCAGGCTGAC | |||||||
KDR | chr4:55972974 | NM_002253 | c.1416A > T | p.Gln472His | F: TACCATGGTAGGCTGCGTTG | 60 | 191 |
R: GGAAGTCCTCCACACTTCTCC | |||||||
MET | chr7:116340262 | NM_001127500 | c.1124A > G | p.Asn375Ser | F: ATTCTTTTCGGGGTGTTCGC | 60 | 201 |
R: TGGGGAACTGATGTGACTTACC | |||||||
PIK3CA | chr3:178927410 | NM_006218 | c.1173A > G | p.Ile391Met | F: AGGTGGAATGAATGGCTGAATTA | 60 | 110 |
R: ACCTCTTTAGCACCCTTTCGG | |||||||
PPP6C | chr9:127912080 | NM_001123355 | c.790C > T | p.Arg264Cys | F: GGTGACAGTATGGTCTGCTCC | 60 | 148 |
R: CGTTGTCGTTCTGGGAGGAA | |||||||
BRAF | chr7:140453136 | NM_004333 | c.1799 T > A | p.Val600Glu | F: GCTTGCTCTGATAGGAAAATGAGAT | 60 | 175 |
R: CATCCACAAAATGGATCCAGACAAC | |||||||
BRAF | chr7:140453136 | NM_004333 | c.1798_1799delGTinsAA | p.Val600Lys | F: GCTTGCTCTGATAGGAAAATGAGAT | 60 | 175 |
R: CATCCACAAAATGGATCCAGACAAC | |||||||
BRAF | chr7:140453135 | NM_004333 | c.1799_1800delTGinsAC | p.Val600Asp | F: GCTTGCTCTGATAGGAAAATGAGAT | 60 | 175 |
R: CATCCACAAAATGGATCCAGACAAC | |||||||
BRAF | chr7:140453145 | NM_004333 | c.1790 T > G | p.Leu597Arg | F: GCTTGCTCTGATAGGAAAATGAGAT | 60 | 175 |
R: CATCCACAAAATGGATCCAGACAAC | |||||||
KIT | chr4:55593464 | NM_000222 | c.1621A > C | p.Met541Leu | F: AGTGGCTGTGGTAGAGATCC | 60 | 427 |
R: CAAAAAGGTGACATGGAAAGC | |||||||
NRAS | chr1:115256529 | NM_002524 | c.182A > G | p.Gln61Arg | F: CACCCCCAGGATTCTTACAG | 60 | 173 |
R: TCCGCAAATGACTTGCTATT | |||||||
NRAS | chr1:115256530 | NM_002524 | c.181C > A | p.Gln61Lys | F: CACCCCCAGGATTCTTACAG | 60 | 173 |
R: TCCGCAAATGACTTGCTATT | |||||||
PTEN | chr10:89720709 | NM_000314 | c.860C > G | p.Ser287Ter | F: GCAACAGATAACTCAGATTGC | 60 | 505 |
R: TTCTTCATCAGCTGTACTCC | |||||||
CDKN2A | chr9:21971089 | NM_001195132 | c.256_268delGCCCGGGAGGGCT | p.Ala86fs | F: AGCTTCCTTTCCGTCATGC | 60 | 0 |
R: GGAAGCTCTCAGGGTACAAAT |