Table 1. Primers, probes and PCR conditions used for real-time PCR and T-RFLP.

Target group	Primer and probe	Sequence (5′–3′)	Reaction system	Thermal profile	Reference
Real-time PCR
Bacteria	Primer Bact1369FB	CGGTGAATACGTTCYCGG	25 µl:12.5 µl Premix Ex Taq™, 1 µl BSA, 0.5 µl each primer and probe, 2 µl template, 8 µl H2O	10 s at 95°C for initial denaturation;35 cycles of 15 s at 95°C, 1 min at 56°C	[55]
	Primer Prok1492R	GGWTACCTTGTTACGACTT
	Probe TM1389F	CTTGTACACACCGCCCGTC
Archaea	Primer Ar364aF	CGGGGYGCASCAGGCGCGAA	25 µl: 12.5 µl of SYBR® Premix Ex Taq™, 1 µl BSA, 0.5 µl each primer, 2 µl template, 8.5 µl H2O	30 s at 94°C for initial denaturation;40 cycles of 20 s at 94°C, 30 s at 59°C,and 30 s at 72°C	[56]
	Primer Ar934b	GTGCTCCCCCGCCAATTCCT
Fungi	Primer NS1	GTAGTCATATGCTTGTCC	25 µl: 12.5 µl of SYBR® Premix Ex Taq™, 1 µl BSA, 0.5 µl each primer, 2 µl template, 8.5 µl H2O	3 min at 95°C for initial denaturation; 40 cycles of 10 s at 95°C, 30 s at 55°C, and 1 min at 72°C	[57]
	Primer FUNG	CATTCCCCGTTACCCGTTG
PCR for T-RFLP
Bacteria	Primer 27F-FAM	GAGTTTGATCCTGGCTCAG	50 µl: 5 µl 10×PCR buffer (MgCl2, 2 mM), 4 µl 2.5 mM dNTPs, 0.5 µl EX-Taq polymerase (5 U µl−1), 1 µl each primer, 1 µl BSA, 4 µl template, 33.5 µl H2O(Reaction system for bacteria, archaea and fungi)	5 min at 94°C; 35 cycles of 45 s at 94°C,45 s at 54°C, 72°C for 90 s; 10 min at 72°C.	[58]
	Primer 1492R	ACGGCTACCTTGTTACGACT
Archaea	Primer Ar364aF	CGGGGYGCASCAGGCGCGAA		35 cycles of 45 s at 94°C, 45 s at 58°C, 60 sat 72°C; 10 min at 72°C.	[56]
	Primer Ar934b-FAM	GTGCTCCCCCGCCAATTCCT
Fungi	Primer NS1-FAM	GTAGTCATATGCTTGTCC		5 min at 94°C; 35 cycles of 30 s at 94°C, 30 sat 56°C, 60 s at 72°C; 10 min at 72°C.	[57]
	Primer FUNG	CATTCCCCGTTACCCGTTG