Mutation spectrum of 122 hemophilia A families from Taiwanese population by LD-PCR, DHPLC, multiplex PCR and evaluating the clinical application of HRM

Lin, Shin-Yu; Su, Yi-Ning; Hung, Chia-Cheng; Tsay, Woei; Chiou, Shyh-Shin; Chang, Chieh-Ting; Ho, Hong-Nerng; Lee, Chien-Nan

doi:10.1186/1471-2350-9-53

Table 1 Primer pairs and PCR conditions for DHPLC analysis.

From: Mutation spectrum of 122 hemophilia A families from Taiwanese population by LD-PCR, DHPLC, multiplex PCR and evaluating the clinical application of HRM

HA Primer pairs and PCR conditions for DHPLC analysis
Exon primer	Sequence (5' to 3')	Length of PCR amplicon (bp)	Annealing Temp. (°C)	DHPLC oven (°C)		Elution profile(B%)
1F	ACATCCAGTGGGTAAAGTTC	356	53	60	55	56–65	52–61
1R	AGACTTACATCCCCACAATC
2F	TTGAAGTGTCCACCAAAATGAACGACT	211	54	59		48–57
2R	GATACCCAATTTCATAAATAGCATTCA
3F	GTACTATCCCCAAGTAACCTT	204	54	59		50–59
3R	CATAGAATGACAGGACAATAGG
4F	TACAGTGGATATAGAAAGGAC	296	54	57		54–63
4R	TGCTTATTTCATCTCAATCCTACGCTT
5F	CCTCCTAGTGACAATTTCCTA	188	54	55		49–58
5R	AGCAGAGGATTTCTTTCAGGAATCCAA
6F	CATGAGACACCATGCTTAGCT	224	54	60		51–60
6R	AACTCTGGTGCTGAATTTGGAAGACCCT
7F	CAGATTCTCTACTTCATAGCCATAG	324	54	58	56	55–64	53–62
7R	ATTAAAAGTAGGACTGGATA
8F	ATATAGCAAGACACTCTGACA	338	54	58		55–64
8R	AGAGAGTACCAATAGTCAAA
9F	AGAGTTGGATTTGAGCCTACC	284	54	58	55	53–62	50–59
9R	CAGACTTTTTCTTCTTACCTGACCTT
10F	GGATTTGATCTTAGATCTCGC	204	53	56	54	50–59	48–57
10R	ATTTTAGTTGTTATTGATGA
11F	TTGAGCTATTTATGGTTTTG	294	53	58.5	56	53–62	51–60
11R	GACATACACTGAGAATGAA
12F	GCATTTCTTTACCCCTTTCA	230	54	59.5	57	51–60	49–58
12R	CTTTATTCACCACCCACTG
13F	TCCTGGGAATAAGATAATGG	393	54	57.5	56	56–65	54–63
13R	AGAGCATACGAATGGCTAGT
14(I)F	ATCTGTGTTATGAGTAACCA	430	54	57.5		57–66
14(I)R	TCATATTTGGCTTCTTGGAG
14(II)F	CATGGGCTATCCTTATCTGA	479	54	56.5		57–66
14(II)R	CATGAACTTTCTTGGCTATT
14(III)F	TCAAAGTTGTTAGAATCAGG	441	54	55.5		56–65
14(III)R	ATTTTGTGCATCTGGTGGAA
14(IV)F	GTCCAACAGAAAAAAGAGGG	481	54	57	54.5	58–67	55–64
14(IV)R	CTACATTTTGCCTAGTGCTC
14(V)F	CTGGCACTAAGAATTTCATG	429	54	57.5	56	57–66	55–64
14(V)R	CCTTCTCATTGTAGTCTATC
14(VI)F	GAAACATTTGACCCCGAGCA	431	54	57.5		56–65
14(VI)R	TTTTGGGCAAGTCTGGTTTC
14(VII)F	CACATACAAGAAAGTTGAGA	436	54	59	57.5	56–65	55–64
14(VII)R	CTCATTTATTGCTGCTATTG
14(VIII)F	GATACCATTTTGTCCCTGAA	415	54	57.5		56–65
14(VIII)R	GTCACAAGAGCAGAGCAAAG
15F	CACCTAGGAAAATGAGGATGT	300	53	58.5	56	53–62	51–60
15R	ATAGTCAGCAAGAAAATAAA
16F	AAGATCCTAGAAGATTATTC	330	50	57.5		53–62
16R	TTAGTACACAAAGACCATTT
17F	TGATGAGAAATCCACTCTGG	349	54	58	56.5	54–63	53–62
17R	GTGCAATCTGCATTTCACAG
18F	GTGGAATCCTCATAGATGTCA	312	53	57		54–63
18R	GAGTAGGTAGAAGAAAGAGCAC
19F	GCAAGCACTTTGCATTTGAG	305	52	59	56	55–64	52–61
19R	AGCAACCATTCCAGAAAGGA
20F	CCATTTTCATTGACTTACATTTGAG	193	53	59.5	56.5	49–58	46–55
20R	AGATATAATCAGCCCAGGTTC
21F	GAATTTAATCTCTGATTTCTCTAC	168	53	60.5	56	48–57	46–55
21R	GAGTGAATGTGAATACATTTCC
22F	AAATAGGTTAAAATAAAGTGTTAT	206	53	48		48–57
22R	GACTAATTACATACCATTAAG
23F	CTCTGTATTCACTTTCCATG	250	54	57		52–61
23R	ACAGTTAGTCACCCTACCCA
24F	GCTCAGTATAACTGAGGCTG	249	54	58.5		51–60
24R	CTCTGAGTCAGTTAAACAGT
25F	AGTGCTGTGGTATGGTTAAG	323	56	58	56.5	55–64	54–63
25R	TTGCTCTGAAAATTTGGTCATA
26F	ATCCTGGACTACTGGAAACA	393	53	63	58.5	56–65	51–60
26R	AGTTAATTCAGGAGGCTTCA

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ISSN: 1471-2350

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