Table 1

Oligonucleotides (A) and conditions (B) for UPQFM-PCR

PCR reaction	Region amplified	Sense primer	Antisense primer	Amount of primer added to volume of 25 μl (pmol)	Product length
Each primer used in PCR 1 reaction begins with 22 bp P2 universal sequence.
A fluorescent label (6-Fam) is attached to the 5` end of the antisense P2 universal primer only.
(A) Oligonucleotides for UPQFM-PCR
PCR 1	Exon 1	E1Fn:	P2-UnivF-tacggccctgaagaagacg	5	321 bp
		E1R:	P2-UnivR-tacctcggtagctgtggatg	5
	Exon 2	E2Fn:	P2-UnivF-agacgaggtttcaccacgtt	2	396 bp
		E2R:	P2-UnivR-gggcttaatttttcaagtggtc	2
	Exon 3	E3Fn:	P2-UnivF-tactgagaccctagtctgtcactg	1,5	364 bp
		E3R:	P2-UnivR-ctaaggaaggaaccagtcctgtat	1,5
	β-globin	BgFn:	P2-UnivF-cacaccctagggttggccaa	2	304 bp
		BgR:	P2-UnivR-acctgtcttgtaaccttgatac	2
PCR 2	All above fragments	P2 Univ F:	tcc gtc tta gct gag tgg cgt a	4
		P2 Univ R:	6-Fam-agg cag aat cga ctc acc gct a	4

PCR reaction	Initial denaturation at 94°C	Each cycle of			No of cycles	Final elongation at 72°C	Template
		Denaturation at 95°C	Annealing at 60°C	Elongation at 72°C
(B) Conditions for UPQFM-PCR
PCR-1	10 minutes	1 minute	2 minutes	1 minute	9 cycles	5 minutes	100 ng genomic DNA
PCR-2	4 minutes	1 minute	2 minutes	1 minute	20 cycles	5 minutes	2 μl PCR 1 product