Active Microbial Communities Inhabit Sulphate-Methane Interphase in Deep Bedrock Fracture Fluids in Olkiluoto, Finland

Table 3

The primers used for amplification of different microbial groups for 454 pyrosequencing. The archaeal 16S rRNA and the mcrA gene transcripts were amplified using a nested PCR approach.

Target	Primer	Sequence	Fragment length (gene location)	Reference
Bacteria 16S rRNA	8F^*; P2^*	5′-AGAGTTTGATCCTGGCTCAG-3′; 5′-ATTACCGCGGCTGCTGG-3′	ca. 500 bp; (V1–V3)	[23]; [24]
Archaea 16S rRNA	A109f; Arch915R	5′-ACKGCTCAGTAACACGT-3′; 5′-GTGCTCCCCCGCCAATTCCT-3′	ca. 800 bp	[25]; [26]
	ARC344f^*; Ar744r^*	5′-ACGGGGCGCAGCAGGCGCGA-3′; 5′-CCCGGGTATCTAATCC-3′	ca. 430 bp; (V3-V4)	[27]; modified from [28]
Methanogens mcrA	mcrA412f; mcr1615r	5′-GAAGTHACHCCNGAAACVATCA-3′; 5′-GGTGDCCNACGTTCATBGC-3′	1.2 kb	[3]; [3]
	ME1^*; ME3r^*	5′-GCMATGCARATHGGWATGTC-3′; TGTGTGAAWCCKACDCCACC-3′	330 bp	[31]; modified from [31]
Sulphate reducer dsrB	2060F^*; dsr4R^*	5′-CAACATCGTYCAYACCCAGGG-3′; 5′-GTGTAGCAGTTACCGCA-3′	370 bp	[29]; [30]

Primers marked with * were equipped with adapter and barcode sequences at the 5′ ends, except if they were used for RT-qPCR. Primers marked with § were used in the qPCR without the adapters and barcodes.