Rumen fermentation and microbial community composition influenced by live Enterococcus faecium supplementation

AMB Express

Table 1 Real time PCR primers used for the quantification of microbial population

Target gene	Primer sequence	Length	Initial denaturation	Denaturation	Annealing	Extension	Cycles	Reference
General bacteria								Denman and McSweeney (2006)
F sequence (5′-3′)	CGGCAACGAGCGCAACCC	130	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
R sequence (5′-3′)	CCATTGTAGCACGTGTGTAGCC	130	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
General anaerobic fungi
F sequence (5′-3′)	GAGGAAGTAAAAGTCGTAACAAGGTTTC	120	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
R sequence (5′-3′)	CAAATTCACAAAGGGTAGGATGATT	120	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
Methanogens
F sequence (5′-3′)	TTCGGTGGATCDCARAGRGC	140	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
R sequence (5′-3′)	GBARGTCGWAWCCGTAGAATCC	140	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
Fibrobacter succinogenes
F sequence (5′-3′)	GTTCGGAATTACTGGGCGTAAA	121	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
R sequence (5′-3′)	CGCCTGCCCCTGAACTATC	121	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
Ruminococcus flavefaciens
F sequence (5′-3′)	CGAACGGAGATAATTTGAGTTTACTTAGG	132	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40
R sequence (5′-3′)	CGGTCTCTGTATGTTATGAGGTATTACC	132	95 °C 2 min	95 °C 15 s	60 °C 60 s	72 °C 30 s	40