Table 1. PCR primers used for PCR, ARISA and real-time PCR of 16S rDNA genes.
| Name | Function | Target group | Sequence (5′—3′) | Amplicon size (bp) | Annealing temp. (°C) | Primer conc. (mℳ) | Ref. |
|---|---|---|---|---|---|---|---|
| A8F | Cloning | Most of Archaea | CGG-TTG-ATC-CTG-CCG-GA | 900 | 57 | 0.4 | Teske et al. (2002) |
| A915R | Sequencing | GTG-CTC-CCC-CGC-CAA-TTC-CT | |||||
| A915F | ARISA | ITS of Archaea | AAA-GGA-ATT-GGC-GGG-GGA-GCA-C | Variable | 55 | 0.4 | Casamayor et al. (2002) |
| A71R (23S) | TCG-GYG-CCG-AGC-CGA-GCC-ATC-C | ||||||
| Mcc495F | Q-PCR | Methanococcales | TAA-GGG-CTG-GGC-AAG-T | 186 | 61 | 1.1 | Yu et al. (2005) |
| McocR | GGA-TCA-ACA-CAT-TTC-ACC-GC | This study | |||||
| MBT857F | Q-PCR | Methanobacteriales | CGW-AGG-GAA-GCT-GTT-AAG-T | 343 | 60 | 0.3 | Yu et al. (2005) |
| MBT1196R | TAC-CGT-CGT-CCA-CTC-CTT | Yu et al. (2005) | |||||
| MmicF | Q-PCR | Methanomicrobiales | GTG-ATA-AGG-GAA-CCY-CGA-G | 126 | 61 | 0.9 | This study |
| MmicR | GCT-ACG-RAC-GCT-TTA-AGC-C | This study | |||||
| MsarF | Q-PCR | Methanosarcinales | GTA-CCT-ACT-AGC-CDA-CRA-CGG | 244 | 64 | 0.9 | This study |
| Msar2R | GCT-ACR-GAC-CCT-TTA-GAC-CC | This study | |||||
| MpyrF | Q-PCR | Methanopyrales | GGC-TAA-TCC-CCG-ATA-GGC | 151 | 61 | 0.8 | This study |
| MpyrR | CTT-GTC-TCA-GTC-CCC-GTC-TC | This study | |||||
| ANME-1F | Q-PCR | ANME-1 | GCT-TTC-AGG-GAA-TAC-TGC | 200 | 60 | 1.1 | Lloyd et al. (2011) |
| ANME-1R | TCG-CAG-TAA-TGC-CAA-CAC | Lloyd et al. (2011) | |||||
| ANME-2aF | Q-PCR | ANME-2a | ACG-GAT-ACG-GGT-TGT-GAG-AG | 151 | 60 | 1.1 | This study |
| ANME-2aR | CTT-GTC-TCA-GTC-CCC-GTC-TC | This study | |||||
| ANME-2cR | Q-PCR | ANME-2c | TCC-TCT-GGG-AAA-TCT-GGT-TG | 224 | 60 | 1.1 | This study |
| ANME-2cF | TCG-TTT-ACG-GCT-GGG-ACT-AC | This study | |||||
| ANME-3F | Q-PCR | ANME-3 | GGA-TTG-GCA-TAA-CAC-CGG | 234 | 60 | 1.1 | This study |
| ANME-3R | TAT-GCT-GGC-ACT-CAG-TGT-CC | This study | |||||
| ARC787F | Q-PCR | Archaea | ATT-AGA-TAC-CCS-BGT-AGT-CC | 273 | 60 | 0.5 | Yu et al. (2005) |
| ARC1059R | GCC-ATG-CAC-CWC-CTC-T | ||||||
| BACT1369F | Q-PCR | Bacteria | CGG-TGA-ATA-CGT-TCY-CGG | 142 | 58 | 0.6 | Suzuki et al. (2000) |
| BACT1492R | GGW-TAC-CTT-GTT-ACG-ACT-T |
Abbreviations: ANME, archaeal anaerobic methanotroph; ARISA, Automated method of Ribosomal Intergenic Spacer Analysis; ITS, Intergenic Transcribed Spacer; Q-PCR, quantitative polymerase chain reaction.