Table 4.

Summary of Laboratory-Developed Methodologies for Sizing of Normal and Premutation FMR1 Alleles

span Lab ID	Primers	Fragment sizing instrument or method	Matrix	Label	Allele sizing calculation	Reference
1	F 5′-6FAM-GGAACAGCGTTGATCACGTGACGTGGTTTC-3′	ABI 310	POP6	6-FAM	(bp-113)/3	10
	R 5′-GGGGCCTGCCCTAGAGCCAAGTACCTTGT-3′
2	F 5′-GACGGAGGCGCCCGTGCCAGG-3′	PAGE and phosphoimaging	6% polyacrylamide (19:1), 7 mol/L urea	32P-dCTP incorporated during PCR	(bp-113)/3	11
	R 5′-TCCTCCATCTTCTCTTCAGCCCT-3′
3	F 5′-6FAM-TGACGGAGGCGCCGCTGCCAGGGGGCGTGC-3′	ABI 377	7% polyacrylamide (19:1), 8.3 mol/L urea	6-FAM	(bp-69)/3	12
	R 5′-GAGAGGTGGGCTGCGGGCGCTCGAGGCCCA-3′
4	F 5′-AGGCGCTCAGCTCCGTTTCGGTTTCACTTC-3′	ABI 3100	POP6	6-FAM	(bp-147)/3	13
	R 5′-6FAM-GTGGGCTGCGGGCGCTCGAGG-3′
5	F 5′-6TETGCTCAGCTCCGTTTCGGTTTCACTTCCGGT-3′	ABI 377	6% polyacrylamide (19:1), 6 mol/L urea	TET	(bp-218)/3	14
	R 5′-AGCCCCGCACTTCCACCACCAGCTCCTCCA-3′
6	F 5′-GCTCAGCTCCGTTTCGGTTTCACTTCCGGT-3′	ABI 3100	POP4	6-FAM	(bp-221)/3)+4	15
	R 5′-6FAM-AGCCCCGCACTTCCACCACCAGCTCCTCCA-3′
7	F 5′-GCTCAGCTCCGTTTCGGTTTCACTTCCGGT-3′	PAGE and autoradiography	6% polyacrylamide (19:1), 7 mol/L urea	32P-dCTP incorporated during PCR	(bp-218)/3	15
	R 5′-AGCCCCGCACTTCCACCACCAGCTCCTCCA-3′
8	F 5′-GACGGAGGCGCCGCTGCCAGG-3′	PAGE and autoradiography	6% polyacrylamide (19:1), 7 mol/L urea	32P-(CGG)5 probe	(bp-62)/3	15
	R 5′-GTGGGCTGCGGGCGCTCGAGG-3′
9	F 5′-6FAM-GTGACGGAGGCGCCGCTGCCA-3′	ABI 3100	POP4	6-FAM	[bp-18)/3]+3	15
	R 5′-AGCTCCTCCATCTTCTCTTCAGCCCTGCTA-3′

R, reverse; F, forward; PAGE, polyacrylamide gel electrophoresis.