TABLE 4.
16S rRNA-targeted probes used for microarray hybridzation.
Original name | Name | Full namea | Primerb | Microarray position(s) | Sequence (5′-3′) | Tm (°C) | ΔG (kcal mol−1) | Specificity | Reference |
---|---|---|---|---|---|---|---|---|---|
CONT-COMP | CTT CCT TCC TTC CTT CCT | Complementary to control oligonucleotide | 35 | ||||||
CONT | A1, A48, B1, B48, C1, C48, D1, D48 | AGG AAG GAA GGA AGG AAG | 51.3 | −18.8 | Control oligonucleotide | 35 | |||
NONSENSE | A2, A25, A47, C25, D47 | AGA GAG AGA GAG AGA GAG | 48.5 | −17.7 | Nonbinding control | 35 | |||
UNIV1390 | UNIV1389a | S-D-Univ-1389-a-A-18 | R | A27, C27, D5 | ACG GGC GGT GTG TAC AAG | 58.3 | −22.2 | Bacteria, not Epsilonproteobacteria | Modified from 51 |
UNIV1390 | UNIV1389b | S-D-Univ-1389-b-A-18 | R | D6 | ACG GGC GGT GTG TAC AAA | 58 | −21.9 | Eucarya | Modified from 51 |
UNIV1390 | UNIV1389c | S-D-Univ-1389-c-A-18 | R | D7 | ACG GGC GGT GTG TGC AAG | 62 | −23.9 | Archaea | Modified from 51 |
EUB338 | EUB338 | S-D-Bact-0338-a-A-18 | A3, A46, C26, D2, D46 | GCT GCC TCC CGT AGG AGT | 59.5 | −22.4 | Most Bacteria | 5 | |
EUB338II | EUB338II | S-*-BactP-0338-a-A-18 | D3 | GCA GCC ACC CGT AGG TGT | 60.7 | −23.0 | Planctomycetes | 14 | |
EUB338III | EUB338III | S-*-BactV-0338-a-A-18 | D4 | GCT GCC ACC CGT AGG TGT | 60.7 | −23.0 | Verrucomirobia | 14 | |
BTWO23a | BTWO663 | S-*-Btwo-0663-a-A-18 | A34, D9 | GGA ATT CCA CCC CCC TCT | 57 | −21.0 | Most Rhodocyclales, but not the Zoogloea and the Azospira lineages | Modified from 4 | |
RHC143 | S-*-RHC-0143-a-A-18 | R | A4, A19, A44, D19 | TCG CTA CGT TAT CCC CCA | 56 | −21.2 | Most members of the Dechloromonas-Ferribacterium-Quadricoccus-Azonexus, the Azospira, and the Rhodocyclus-Propionivibrio-Accumulibacter lineages, Zoogloea spp., and few members of the Azoarcus-Thauera-Denitromonas lineage | This study | |
S-G-Rhc-0175-a-A-18 | RHC175a | S-*-RHC-0175-a-A-18 | A42, D17 | TGC TCA CAG AAT ATG CGG | 53 | −20.0 | Most members of the Dechloromonas-Ferribacterium-Quadricoccus-Azonexus, the Azospira, and the Rhodocyclus-Propionivibrio-Accumulibacter lineages | 25 | |
S-G-Rhc-0439-a-A-18 | RHC439 | S-*-RHC-0439-a-A-18 | A43, D18 | CNA TTT CTT CCC CGC CGA | 54.9-58.3 | Rhodocyclus spp., most members of the Candidatus “Accumulibacter” cluster, Azospira lineage | 25 | ||
RHAC855 | S-*-RhAc-0855-a-A-18 | A7 | TCA CGC GTT AGC TAC GGC | 58.2 | −22.8 | Rhodocyclus spp., most members of the Candidatus “Accumulibacter” cluster | This study | ||
PAO846 | ACCBA846 | S-G-Accba-0846-a-A-18 | A10 | AGC TAC GGC ACT AAA AGG | 52.6 | −19.7 | Most members of the Candidatus “Accumulibacter” cluster | Modified from 13 | |
PAO651 | ACCBA651 | S-G-Accba-0651-a-A-18 | A6 | CCC TCT GCC AAA CTC CAG | 55.8 | −20.9 | Most members of the Candidatus “Accumulibacter” cluster | 13 | |
ACCBA443 | S-*-Accba-0443-a-A-18 | A8 | CAA GCA ATT TCT TCC CCG | 52.2 | −19.6 | Some members of the Candidatus “Accumulibacter” cluster | This study | ||
S-G-Rhx-0456-a-A-17 | ACCBA455 | S-*-Accba-0455-a-A-18 | A9 | AGG GTA TTA ACC CAA GCA | 51.4 | −18.8 | Some members of the Candidatus “Accumulibacter” cluster | 25 | |
RHO828 | S-G-Rho-0828-a-A-18 | A12 | TTA ACC CCA CCA ACA CCT | 54.4 | −20.0 | Rhodocyclus spp. | This study | ||
RHO842 | S-G-Rho-0842-a-A-18 | A13 | CGG CAC TAA TGG GTT TAA | 50.5 | −18.8 | Rhodocyclus spp. | This study | ||
RHOTE1280 | S-S-Rho.te-1280-a-A-18 | A15 | CGA TCG GCT TTG CGG GAT | 58.9 | −22.6 | Rhodocyclus tenuis | This study | ||
AZP471 | S-G-Azp-0471-a-A-18 | A29 | GTA CCG TCA TCA ACA ACG | 51.4 | −19.5 | Most Azospira spp. | This study | ||
AZP456 | S-G-Azp-0456-a-A-18 | A30 | ACG GAT ATT AGC CGT TGC | 53.1 | −20.1 | Most Azospira spp. | This study | ||
AZP737 | S-G-Azp-0737-a-A-18 | A28 | GTC AGT ACT AAC CCA GGG | 51.6 | −19.1 | Most Azospira spp. | This study | ||
DAF1030 | S-*-D.A.F-1030-a-A-18 | A22, A45, D20 | TGT GTT CCA GCT CCC TTT | 54.7 | −20.2 | Some bacteria of the Dechloromonas-Ferribacterium-Quadricoccus-Azonexus lineage | This study | ||
DEMFE455 | S-*-DemFe-0455-a-A-18 | A21 | AGG GTA TTA ACC CAT GCG | 52.5 | −19.5 | Ferribacterium limneticum, few Dechloromonas spp. | This study | ||
QUACO135 | S-*-Quaco-0135-a-A-18 | D40 | TTA TCC CCC ACT CAA TGG | 51.8 | −19.0 | Quadricoccus australiensis, reactor clone PHOS-HE23 | This study | ||
DCMAG455 | S-S-Dcm.ag-0455-a-A-18 | A20 | CAG GTA TTA GCT GAT GCG | 50.7 | −19.1 | Dechloromonas agitata | This study | ||
A08KA458 | S-*-A08KA-0459-a-A-18 | D44 | ACA CCC CGT ATT AGA GAG | 50.7 | −18.7 | Oral strain A08KA lineage | This study | ||
TH14 | RHC175b | S-*-RHC-0175-b-A-18 | A39, C4, C28, C40, D14 | CCC TCA GGA CGT ATG CGG | 57.9 | −22.1 | Some Thauera, Azoarcus, Zoogloea, Sterolibacterium, and Azovibrio spp. | Modified from 26 | |
TH3 | RHC222 | S-*-RHC-0222-a-A-18 | A36, B2, C2, C41, D11 | ACA TCG GCC GCT CCA ATC | 58.6 | −22.4 | Some members of the Azoarcus-Thauera-Denitromonas, Zoogloea, and Azovibrio lineages | 26 | |
AZV211 | S-G-Azv-0211-a-A-18 | C42 | TCC AAT CGC ACA AGG TCC | 55.7 | −21.2 | Azovibrio spp. | This study | ||
AZVRE847 | S-*-Azv.re-0847-a-A-18 | C43 | TAG CTC CGT TAC TAA TAG | 45.1 | −17.1 | Azovibrio restrictus | This study | ||
AT1458 | ATD1459 | S-*-ATD-1459-a-A-18 | A,R,Z | A35, D10 | TCT CAC CGT GGT AAG CGC | 58 | −22.4 | Most members of the Azoarcus-Thauera-Denitromonas lineage | Modified from 43 |
RHC630 | S-*-RHC-0630-a-A-18 | A40, D15, D25 | TGC AGT CAC AAA CGC AGT | 56.1 | −21.2 | Most Thauera, Zoogloea, and Rhodocyclus spp. | This study | ||
AZA1006 | S-*-Aza-1006-a-A-18 | A38, B4, D13 | TCC CTG ATC TCT CAA GGA | 52.1 | −19.5 | Most members of the Azoarcus cluster | This study | ||
AZA483 | S-*-Aza-0483-a-A-18 | B6 | CTT CTT CTG ACA GTA CCG | 49.7 | −18.6 | Azoarcus cluster | This study | ||
Azo644 | AZA645 | S-*-Aza-0645-a-A-18 | B5, B43 | GCC GTA CTC TAG CCG TGC | 58.3 | −22.5 | Most members of the Azoarcus cluster | 24 | |
Azo1251 | AZA1252 | S-*-Aza-1252-a-A-18 | B7 | TCG CGC TTT GGC AGC CCT | 64.1 | −24.8 | Azoarcus evansii, Azoarcus toluvorans, Azoarcus tolulyticus, Azoarcus toluclasticus, and related Azoarcus spp. | Modified from 24 | |
AZA444 | S-*-Aza-0444-a-A-18 | B9 | GGA AGC GTT TTC TTT CCG | 52.4 | −19.9 | Azoarcus evansii, Azoarcus tolulyticus str. Td-3, Azoarcus tolulyticus str.Td-19, Azoarcus sp. ToN1 | This study | ||
AZTOLY452 | S-S-Az.toly-0452-a-A-18 | B10 | GTA TTG ACC CAC CCG ATT | 52.4 | −19.5 | Azoarcus tolulyticus | This study | ||
AZANBU228 | S-*-Az.an.bu-0228-a-A-18 | B12 | AAT CCG ACA TCA GCC GCT | 57.7 | −21.8 | Azoarcus anaerobius, A. buckelli, and related Azoarcus spp. | This study | ||
AZA844 | S-*-Aza-0844-a-A-18 | B20 | TGC GTC ACT CAG CGC GTT | 61.1 | −23.7 | Azoarcus spp. PH002 and CR23 | This study | ||
AZA452 | S-*-Aza-0452-a-A-18 | B19 | CTA TTC ACG CAC CCG ATT | 53 | −20.0 | Azoarcus spp. PH002 and CR23 | This study | ||
AZA463 | S-*-Aza-0463-a-A-18 | B15 | ATC CAG GCA CGC TAT TCA | 54.6 | −20.3 | Azoarcus spp. PbN1 and HxN1 | This study | ||
AZA835 | S-*-Aza-0835-a-A-18 | B16 | CAG AAA GTT ACC TTC CCG | 50.5 | −18.8 | Azoarcus sp. PbN1 | This study | ||
AZAN465 | S-S-Az.an-0465-a-A-18 | B14 | TCA TCC AGG CTC GCT ATT | 54 | −20.3 | Azoarcus anaerobius | This study | ||
AZAN130 | S-S-Az.an-0130-a-A-18 | R | B13 | CCC CTC GAC TGG GTA CGT | 59 | −22.2 | Azoarcus anaerobius | This study | |
S-*-OTU2-0132-a-A-18 | ATDe132 | S-*-ATDe-0132-a-A-18 | R | B24 | CCC CCA CAA CAT GGG TAC | 55.9 | −20.9 | Activated sludge clones A33, H25, H30, H35, S3, S10, and S23 of the Azoarcus-Thauera-Denitromonas lineage | 27 |
Original name | Name | Full namea | Primerb | Microarray position(s) | Sequence (5′-3′) | Tm (°C) | ΔG (kcal mol−1) | Specificity | Reference |
ATDe442 | S-*-ATDe-0442-a-A-18 | B25 | ACC CCG TTT CTT CCC AAC | 55.7 | −20.8 | Activated sludge clones A33, H25, H30, H35, S3, S10, and S23 of the Azoarcus-Thauera-Denitromonas lineage | This study | ||
ATDe830 | S-*-ATDe-0830-a-A-18 | B26 | CGT TAC CGC TCC GAA CAA | 55.8 | −21.4 | Activated sludge clones A33, H25, H30, H35, S3, S10, and S23 of the Azoarcus-Thauera-Denitromonas lineage | This study | ||
S-*-OTU2-0467-a-A-20 | ATDe467 | S-*-ATDe-0467-a-A-18 | B27 | CGT CAT TAG GAT CCT ATG | 46.6 | −17.2 | Activated sludge clones A33, H25, H30, H35, S3, S10, and S23 of the Azoarcus-Thauera-Denitromonas lineage | Modified from 27 | |
S3-486 | S-*-S3-0486-a-A-18 | B31 | GTG CTT CTT CCG TCG GTA | 54.9 | −21.4 | Activated sludge clone S3 | This study | ||
A33-587 | S-*-A33-0587-a-A-18 | B34 | CAC CTG TCT TAC CAA ACC | 50.7 | −18.9 | Activated sludge clone A33 | This study | ||
DENAR176 | S-S-Denar-0176-a-A-18 | C16 | TCC CTC AGG AAA TAT GCG | 52.1 | −19.6 | Denitromonas aromaticus | This study | ||
DENAR453 | S-S-Denar-0453-a-A-18 | C17 | CGT ATT CGG GGC GAT GAT | 55.3 | −21.0 | Denitromonas aromaticus | This study | ||
DENAR845 | S-S-Denar-0845-a-A-18 | C18 | GCT GCG TTA CCC AGA AAG | 54.2 | −20.6 | Denitromonas aromaticus | This study | ||
AZCOM447 | S-S-Az.com-0447-a-A-18 | B42 | AGC CCA CAC GTT TTC TTC | 53.8 | −20.2 | Azoarcus communis | This study | ||
TH5 | AZIND1023 | S-S-Az.ind-1023-a-A-18 | A37, B3, B38, C3, D12 | CTG GTT CCC GAA GGC ACC | 58.9 | −22.3 | Azoarcus indigens, Azoarcus sp. BH72 | Modified from 26 | |
AZIND433 | S-S-Az.ind-0433-a-A-18 | B39 | CTT TCC GTC CGA AAG AGC | 53.8 | −20.5 | Azoarcus indigens, Azoarcus sp. BH72 | This study | ||
AZIND449 | S-St-Az.ind-0449-a-A-18 | B40 | TTA GCC CGC GCG ATT TCT | 58.2 | −22.2 | Azoarcus sp. BH72 | This study | ||
AZIND455 | S-St-Az.ind-0455-a-A-18 | B41 | CGG GTA TTG GCC GAA GCG | 59.4 | −23.0 | Azoarcus indigens (T) | This study | ||
THAU832 | S-G-Thau-0832-a-A-18 | C5 | TGC ATT GCT GCT CCG AAC | 57 | −21.7 | Thauera spp. | This study | ||
THAU455a | S-*-Thau-0455-a-A-18 | C6 | ACT ATG TTA GAG TGC GCG | 52.6 | −19.9 | Thauera chlorobenzoica, Thauera mechernichensis | This study | ||
THAU443 | S-*-Thau-0443-a-A-18 | C7 | AAC ACG ATT TCT TCC CGG | 53.2 | −20.0 | Thauera selenatis, Thauera phenylacetica, and related Thauera spp. | This study | ||
THAU580 | S-*-Thau-0580-a-A-18 | C8 | CTT ACA AAA CCG GCC TCG | 54.2 | −20.6 | Thauera selenatis, soil clone AX39 | This study | ||
THAU455b | S-*-Thau-0455-b-A-18 | C9 | ACT ATG TTA GAG TCG CCG | 51.6 | −19.4 | Thauera sp. PIV-1, TCB-transforming microbial consortium clone SJA-186 | This study | ||
THAU468 | S-*-Thau-0468-a-A-18 | C10 | CCG TCA TCC AGC GAC TAT | 54.5 | −20.6 | Thauera sp. PIV-1, TCB-transforming microbial consortium clone SJA-186 | This study | ||
THATE461 | S-S-Tha.te-0461-a-A-18 | C11 | CCA CAC CCT ATG TTA GAG | 49.2 | −18.2 | Thauera terpenica | This study | ||
ZOGLO828 | S-G-Zoglo-0828-a-A-18 | C30 | TCT CCT CAC CGA ACA ACT | 53.6 | −20.1 | Zoogloea spp. | This study | ||
S-*-OTU1-1415-a-A-20 | ZOGLO1416 | S-G-Zoglo-1416-a-A-18 | C29 | TCT GGT AAA CCC CAC TCC | 54.1 | −20.2 | Zoogloea spp. | Modified from 27 | |
ZRA (ZRA23a) | ZOGLO647 | S-*-Zoglo-0647-a-A-18 | C31 | CTG CCG TAC TCT AGT TAT | 48.2 | −17.8 | Most members of the Zoogloea lineage, not Z. resiniphila | 45 | |
ZOGLO455 | S-*-Zoglo-0455-a-A-18 | C32 | AGA GTA TTA TCC TGC GCG | 52.1 | −19.6 | Some members of the Zoogloea lineage (activated sludge clones H13, H11, H22, H10, H40, H27, and S21), not Z. ramigera and Z. resiniphila | This study | ||
S-*-H7-1014-a-A-18 | H7-1014 | S-*-H7-1014-a-A-18 | C33 | TCG GGC ACC CCT CAA TCT | 59.4 | −22.3 | Activated sludge clone H7 | 27 | |
ZORAM211 | S-S-Zo.ram-0211-a-A-18 | C34 | TCG TAT AAC GTG AGG CCT | 53.4 | −20.2 | Zoogloea ramigera | This study | ||
ZORAM441 | S-S-Zo.ram-0441-a-A-18 | C35 | TGC GAT TTC TTT CCA CCT | 52.6 | −19.5 | Zoogloea ramigera | This study | ||
S-*-OTU3-1426-a-A-18 | STEBA1426 | S-*-Steba-1426-a-A-18 | A, R, Z | D26 | ACT ACC TAC TTC TGG TGG | 50.6 | −18.5 | Some members of the Sterolibacterium lineage | 27 |
STEBA468 | S-*-Steba-0468-a-A-18 | D28 | CCG TCA TTA GTA GCC CGT | 54.5 | −20.5 | Some members of the Sterolibacterium lineage (activated sludge clones S28, A13, S40, H12, H23, and H20) | This study | ||
S-*-OTU3-0445-a-A-20 | STEBA448 | S-*-Steba-0448-a-A-18 | D27 | TAG GGG CCA CCG TTT CGT | 59.8 | −23.1 | Some members of the Sterolibacterium lineage (activated sludge clones S28, A13, S40, H12, H23, and H20) | Modified from 27 | |
STEBA635 | S-*-Steba-0635-a-A-18 | D29 | AGT CCT ACA GTC ACA AAC | 49.3 | −18.2 | Few members of the Sterolibacterium lineage (activated sludge clones S28 and A13) | This study | ||
STEBA643 | S-*-Steba-0643-a-A-18 | D30 | CAC ACT CGA GTT ATG CAG | 50.8 | −19.2 | Few members of the Sterolibacterium lineage (activated sludge clones S40, H12, H23, and H20) | This study | ||
STEBA214 | S-*-Steba-0214-a-A-18 | D32 | CGC TCC TCT CGC GCG AGG | 63.5 | −25.1 | Few members of the Sterolibacterium lineage (clones SBR1001, SBR2080, and GC24) | This study | ||
BONE23a | BONE663c | S-*-Bone-0663-a-A-18 | A33, D8 | GGA ATT CCA TCC CCC TCT | 54.1 | −19.9 | Beta1 group of Betaproteobacteria | Modified from 4 | |
RHOTE206c | S-S-Rho.te-0206-a-A-18 | A14 | AAG CGC AAG GTC CTA AGA | 54.4 | −20.5 | Rhodocyclus tenuis | This study | ||
PPV1239c | S-*-Ppv-1239-a-A-18 | A11 | ACC CTC TGA ACC GAC CAT | 56.4 | −20.9 | Propionivibrio spp., few members of the Candidatus “Accumulibacter” cluster | This study | ||
AZA1269c | S-*-Aza-1269-a-A-18 | B8 | AAG GGA TTG GCT CCA GCT | 57.2 | −21.1 | Azoarcus evansii, Azoarcus toluvorans, Azoarcus tolulyticus, Azoarcus toluclasticus, and related Azoarcus spp. | This study | ||
AZA829c | S-*-Aza-0829-a-A-18 | B18 | GTT ACC GCA CCG AAC AAC | 54.7 | −21.0 | Azoarcus spp. EbN1 and pCyN1 | This study | ||
AZA234c | S-*-Aza-0234-a-A-18 | B17 | CCA GCT AAT CCG ACA TCA | 52.1 | −19.4 | Azoarcus spp. EbN1 and pCyN1 | This study | ||
AZA221c | S-S-Aza-0221-a-A18 | B45 | CAT CGG CCA CTC CAA TCA | 55.5 | −20.9 | Azoarcus sp. LU1 | This study |
Name of 16S rRNA gene-targeted oligonucleotide probe based on the nomenclature of Alm et al. (2).
When “Rhodocyclales” subgroup-selective primer pair A, R, or Z was used for microarray analysis, this probe has its target site outside the amplified 16S rRNA gene fragments and must be ignored during interpretation of the hybridization pattern.
Probe was removed from the RHC-PhyloChip because it hybridized nonspecifically to many reference organisms that have mismatches in the 16S rRNA gene target site.