Table 4.

Primers used to generate PCR products containing Bifidobacterium species-specific primer-binding sites (qPCR standards)^a

Target species	Primerb	Sequence (5′ to 3′)	PCR conditions	Amplicon
				Size (bp)	Molecular mass (g/mol)c
B. adolescentis	B_ado-std-f	GATCAGATCGCTGCCACCGCAAC	94°C, 3 min; 30 times (94°C, 30 s; 69°C, 30 s; 72°C, 30 s); 72°C, 10 min	320	197,799.00
	B_ado-std-r	ATCAGCAGCGGCTTGCCGGTCTT
B. angulatum	B_ang-std-f	AGTGCTCGAAGACCCGTACATCCT	94°C, 3 min; 30 times (94°C, 30 s; 65°C, 30 s; 72°C, 30 s); 72°C, 10 min	164	101,387.60
	B_ang-std-r	GGATGTTGTTCAGGATCAGGGTCG
B. animalis	B_ani-std-f	GTCACCGTCGAAGACAACAACC	94°C, 3 min; 30 times (94°C, 30 s, 65°C, 30 s; 72°C, 30 s); 72°C, 10 min	299	184,812.60
	B_ani-std-r	CAGGACTTGAAGGTGCCACGG
B. bifidum	B_bif-std-f	CAAGGACGTGGAGACCAAG	94°C, 3 min; 30 times (94°C, 30 s; 65°C, 30 s; 72°C, 30 s); 72°C, 10 min	391	241,687.40
	B_bif-std-r	CTTGTTCAGGATGAGGGTCG
B. breve	B_bre-std-f	CCACCGAGGTCATCGTC	94°C, 3 min; 30 times (94°C, 20 s; 58°C, 20 s; 72°C, 20 s); 72°C, 10 min	322	199,032.80
	B_bre-std-r	TGCTGGGAGGAGACCTTG
B. catenulatum	B_cat-std-f	GTCGTGGTATCGAGAAG	94°C, 3 min; 30 times (94°C, 30 s; 47°C, 30 s; 72°C, 30 s); 72°C, 10 min	244	150,819.60
	B_cat-std-r	TAGCCCTTGTCGAAACG
B. dentium	B_den-std-f	CCACTACCGCAACTGTGCTG	94°C, 3 min; 30 times (94°C, 50 s; 62°C, 50 s; 72°C, 50 s); 72°C, 10 min	400	247,244.00
	B_den-std-r	GTCAGGAGGATGTACGGGTC
B. gallicum	B_gal-std-f	TGAGAAGGCTTCCGACG	94°C, 3 min; 30 times (94°C, 30 s; 57°C, 30 s; 72°C, 30 s); 72°C, 10 min	242	149,590.80
	B_gal-std-r	GGGAGATGTAGCCCTTG
B. longum	B_lon-std-f	CTGAGGCTCTGGACAAGGTCG	94°C, 3 min; 30 times (94°C, 30 s; 66°C, 30 s; 72°C, 30 s); 72°C, 10 min	323	199,651.20
	B_lon-std-r	GGTGCCACGGATGTTGTTCAGG
B. pseudocatenulatum	B_pcat-std-f	CGTCGTGGTATCGAGAAGGCTTCCG	94°C, 3 min; 30 times (94°C, 50 s; 64°C, 50 s; 72°C, 50 s); 72°C, 10 min	434	268,244.60
	B_pcat-std-r	AAGGTCGGCAGAGCCTCGCCAT
B. pseudolongum	B_plon-std-f	CCTCAAGAACGTTGTGGC	94°C, 3 min; 30 times (94°C, 30 s; 49°C, 30 s; 72°C, 30 s); 72°C, 10 min	420	259,607.00
	B_plon-std-r	CCGGTCTTCATCACGAG
B. thermophilum	B_the-std-f	GAAGGCCTGAAGAACGTGGTCG	94°C, 3 min; 30 times (94°C, 40 s; 65°C, 40 s; 72°C, 40 s); 72°C, 10 min	449	277,522.60
	B_the-std-r	TCAGCCACGATCAGCAGCGGA

^aThe target sites are located at least 17 bp up- and downstream of the Bifidobacterium species-specific primer-binding sites to prevent their degradation during storage.

^bf, forward primer; r, reverse primer.

^cRefers to the double-stranded amplicon.