Development of a Facile Fluorescent Assay for the Detection of 80 Mutations Within the p53 Gene

Hodgson, Darren R.; Foy, Carole A.; Partridge, Maxine; Pateromichelakis, Stelios; Gibson, Neil J.

doi:10.1007/BF03402148

Molecular Medicine

Table 1 Sequences of ARMS primers used to detect 80 mutations in the p53 gene

From: Development of a Facile Fluorescent Assay for the Detection of 80 Mutations Within the p53 Gene

Well	Codon	Change	Frequency	ARMS Primer (5′ to 3′)	Reverse PCR Primer/Molecular Beacon
3	132	AAG to AGG	0.357	ggt ctt gac cag ttg gca aaa cat tc	861-98/5aU
4	132	AAG to CAG	0.071	tcc tac agt act ccc ctg ccc tca tcc	862-98/6aL
5	135	TGC to TAC	0.071	gta ctc ccc tgc cct cac caa gat gtt gta	862-98/6aL
6	141	TGC to TAC	0.143	aac aag atg ttt tgc caa ctg gcc aag aca ta	862-98/6aL
7	151	CCC to TCC	0.214	tgg cgt gga tgc ggg tgc cgg gcg gag a	861-98/5aU
8	151	CCC to ACC	0	cct gtg cag ctg tcg gtt gat tcc ata a	862-98/6aL
9	152	CCG to CTG	0.143	gca gct gtg ggt tga ttc cac acc tct	862-98/6aL
10	156	CGC to CCC	0.071	cca cac ccc cgc ccg gca ctc c	862-98/6aL
11	157	GTC to TTC	0.929	aca ccc ccg ccc ggc acc ctc t	862-98/6aL
12	158	CGC to CTC	0.286	cac acc ccc tcc cgg cac ccg cgt gct	862-98/6aL
15	158	CGC to CAC	0.571	gat tcc aca ccc ccg ccc ggc acc ctc gta ca	862-98/6aL
16	161	GCC to ACC	0.143	gcc agg cac ccg cgt ccg cgc cag ga	862-98/6aL
17	163	TAC to TGC	0.429	tcc gtc atg tgc tgt gac tgc tgg c	861-98/5aU
18	173	GTG to ATG	0.143	cta caa gca gtc aca gca cat gac gga gga ta	862-98/6aL
19	173	GTG to TTG	0.357	aca agc agt cac agc aca tgc cgg agg att	862-98/6aL
20	175	CGC to CAC	5.071	cac agc aca tga cgg agg ttg tga ggg a	862-98/6aL
21	175	CGC to AGC	0	cac agc aca tga cgg agg ttg tga cga	862-98/6aL
22	175	CGC to TGC	0.071	cac agc aca tga cgg agg ttg tga agt	862-98/6aL
23	175	CGC to CCC	0	aca gca cat gac gga ggt tgt gag ccc	862-98/6aL
24	175	CGC to CTC	0	aca gca cat gac gga ggt tgt gag tct	862-98/6aL
27	176	TGC to TAC	0.286	cat gac gga ggt tgt gag gcg gta	862-98/6aL
28	176	TGC to TTC	4.214	gca cat gac gga ggt tgt gag gcg gtt	862-98/6aL
29	177	CCC to CGC	0	cat gac gga ggt tgt gag gcg ctg ctg	862-98/6aL
30	179	CAT to CGT	0.429	agg ttg tga ggc gct gcc ccc act g	862-98/6aL
31	179	CAT to TAT	0.500	tga cgg agg ttg tga ggc gct gcc ccc att	899-97/6aL
32	192	CAG to TAG	0.214	gat tgc tct tag gtc tgg ccc ctc ttt	694-98/6bL
33	193	CAT to CGT	0.286	gct ctt agg tct ggc ccc tcc tca ccg	694-98/6bL
34	195	ATC to ACC	0.500	ctt agg tct ggc ccc tcc tca gca tct gac	694-98/6bL
35	196	CGA to TGA	1.000	ata ctc cac acg caa att tcc ttc cac aca	861-98/5aU
36	205	TAT to TGT	1.000	cga aaa gtg ttt ctg tca tcc aat c	863-98/6aL
39	213	CGA to TGA	0.786	gga gta ttt gga tga cag aaa cac ttc tt	694-98/6bL
40	220	TAT to TGT	1.571	aca tag tgt ggt ggt gcc gtg	694-98/6bL
41	228	GAC to AAC	0	gcc tgt gtt atc tcc tag gtt ggc tgt a	644-98/7aL
42	234	TAC to TGC	0.714	ccc atg cag gaa ctg tta cac atg tag atc c	643-98/7bU
43	237	ATG to ATA	0.571	gac tgt acc acc atc cac tac aac tac aaa	644-98/7aL
44	238	TGT to TAT	0.214	ctg tac cac cat cca cta caa cta cat cta	644-98/7aL
45	241	TCC to TTC	0.214	tcc agt tca tgc cgc cca tgc aca	643-98/7bU
46	242	TGC to TTC	0.500	acc atc cac tac aac tac atg tgt aac agt tct tt	644-98/7aL
47	244	GGC to TGC	0.214	cta caa cta cat gtg taa cag ttc ctg cat ct	644-98/7aL
48	245	GGC to AGC	1.714	tac gtg tgt aac agt tcc tgc atg gtc a	644-98/7aL
51	245	GGC to GAC	0.786	cat gtg taa cag ttc ctg cat ggg cta	644-98/7aL
52	245	GGC to TGC	0.429	act aca tgt gta aca gtt cct gca tgg tct	644-98/7aL
53	245	GGC to GTC	0.214	cat gtg taa cag ttc ctg cat ggg agt	644-98/7aL
54	245	GGC to CGC	0	aca tgt gta aca gtt cct gca tgg ccc	644-98/7aL
55	245	GGC to GCC	0	cat gtg taa cag ttc ctg cat ggg agc	644-98/7aL
56	248	CGG to CAG	2.571	aca gtt cct gca tgg gcg gca tga aac a	644-98/7aL
57	248	CGG to TGG	2.071	aac agt tcc tgc atg ggc ggc atg aag t	644-98/7aL
58	248	CGG to CTG	0.357	taa cag ttc ctg cat ggg cgg tat gaa tct	644-98/7aL
59	248	CGG to GGG	0.143	gta aca gtt cct gca tgg gcg gcg tga agg	644-98/7aL
60	248	CGG to CCG	0.071	tgt aac agt tcc tgc atg ggc ggc atg aac ac	644-98/7aL
63	249	AGG to AGT	0.357	ttc ctg cat ggg cag cat gaa ccg gtg t	644-98/7aL
64	249	AGG to ATG	0	ttc ctg cat ggg cga cat gaa ccg tat	644-98/7aL
65	249	AGG to AGC	0	gca tgg gcg gca tga acc gga ac	644-98/7aL
66	249	AGG to GGG	0.286	ttc gag tgt gat gat ggt gag gat ggg cgc	643-98/7bU
67	249	AGG to TGG	0.214	ttg cag tgt gat gat ggt gag gat ggg aca	643-98/7bU
68	249	AGG to ACG	0.071	aac agt tcc tgc atg ggc gac atg aac cgg tc	644-98/7aL
69	249	AGG to AAG	0	atg ggc agc atg aac cga aa	644-98/7aL
70	258	GAA to AAA	0	ggc cca tcc tca cca tca tca cac tta	644-98/7aL
71	258	GAA to GGA	0.071	cca tcc tca cca tca tca cac tgc g	644-98/7aL
72	266	GGA to GTA	0	tcc tat cct gag tag tgg taa tct act tgt	629-98/8aL
75	266	GGA to AGA	0.286	ttc cta tcc tga gta gtg gta atc tac gga	629-98/8aL
76	266	GGA to GAA	0.214	tcc tat cct gag tag tgg taa tct act gaa	629-98/8aL
77	272	GTG to ATG	0	atc tac tgg gac gga aca gct ttg tga	629-98/8aL
78	273	CGT to CAT	2.429	ggg acg gaa cag ctt tga ggt gaa	629-98/8aL
79	273	CGT to TGT	1.143	act ggg acg gaa cag ctt tga ggt ct	629-98/8aL
80	273	CGT to CTT	0.714	tac tgg gac gga aca gct ttg agg tac t	629-98/8aL
81	273	CGT to AGT	0	tac tgg gac gga aca gct ttg agg aga	629-98/8aL
82	273	CGT to GGT	0.143	tac tgg gac gga aca gct ttg agg agg	629-98/8aL
83	273	CGT to CCT	0.071	act ggg acg gaa cag ctt tga ggt gac	629-98/8aL
84	275	TGT to TAT	0.357	acg gaa cag ctt tga ggt gcg tgt tga	629-98/8aL
87	278	CCT to CTT	0.214	aac agc ttt gag gtg cgt gtt tgt gcc tgt gt	629-98/8aL
88	278	CCT to TCT	0.786	gaa cag ctt tga ggt gcg tgt ttg tgc cta tt	629-98/8aL
89	280	AGA to ACA	0.071	ggt gcg tgt ttg tgc ctg tcc tgg aac	629-98/8aL
90	282	CGG to TGG	1.643	tgc gtg ttt gtg cct gtc ctg gga gag agt	629-98/8aL
91	282	CGG to CCG	0	gtg cgt gtt tgt gcc tgt cct ggg aga gac gc	629-98/8aL
92	282	CGG to GGG	0.071	ggt gcg tgt ttg tgc ctg tcc tgg gag agt cg	629-98/8aL
93	282	CGG to CAG	0	gtg cgt gtt tgt gcc tgt cct ggg aga gac ga	629-98/8aL
94	285	GAG to AAG	0.571	gcc tgt cct ggg aga gac cgg cgc aga a	629-98/8aL
95	286	GAA to AAA	0.214	tcc tgg gag aga ccg gcg cac agc ga	629-98/8aL
96	298	GAG to TAG	1.000	ccg caa gaa agg gga gcc tca cca gt	629-98/8aL

Key to legend:
Well = position on assay plate,
codon = p53 codon
change = mutation in p53 gene sequence
Frequency = frequency of mutation occurance in p53 database [34] as a percentage of all mutations reported for head and neck cancer (excluding data from cell lines)
ARMS primer = primer sequence used to detect mutation
Reverse PCR Primer/Molecular beacon = reverse PCR primer used with ARMS primer and molecular beacon used to detect PCR amplification (see table 3 for sequences).

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ISSN: 1528-3658

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