ImProm-II 42°C : SuperScriptIV ImProm-II 55°C : TGIRT Phylum Class Phylum Class 0.64 Acidobacteria Acidobacteriia Acidobacteria Acidobacteriia Blastocatellia Blastocatellia Holophagae Holophagae Subgroup Subgroup Thermoanaerobaculia 0.62 Thermoanaerobaculia Actinobacteria MB-A2-108 Actinobacteria MB-A2-108 0.62 Rubrobacteria Rubrobacteria Nitriliruptoria Nitriliruptoria 0319-7L14 0319-7L14 Thermoleophilia Thermoleophilia Acidimicrobiia 0.60 Acidimicrobiia Actinobacteria Actinobacteria Armatimonadetes uncultured Armatimonadetes uncultured 0.60 Fimbriimonadia Fimbriimonadia Bacteroidetes Bacteroidia Bacteroidetes Bacteroidia Chloroflexi Dehalococcoidia Chloroflexi Dehalococcoidia Ktedonobacteria 0.58 Ktedonobacteria TK10 TK10 KD4-96 KD4-96 0.58 JG30-KF-CM66 JG30-KF-CM66 Gitt-GS-136 Gitt-GS-136 Anaerolineae Anaerolineae Chloroflexia 0.56 Chloroflexia Cyanobacteria Oxyphotobacteria Cyanobacteria Oxyphotobacteria Deinococcus-Thermus Deinococci Deinococcus-Thermus Deinococci Entotheonellaeota Entotheonellia Entotheonellaeota Entotheonellia 0.56 Fibrobacteres Fibrobacteria Fibrobacteres Fibrobacteria Firmicutes Bacilli Firmicutes Bacilli Nitrospira Nitrospira Gemmatimonadetes S0134 0.54 Gemmatimonadetes S0134 BD2-11 BD2-11 Gemmatimonadetes Gemmatimonadetes 0.54 Longimicrobia Longimicrobia Patescibacteria Saccharimonadia Patescibacteria Saccharimonadia Phycisphaerae Phycisphaerae Planctomycetacia 0.52 Planctomycetacia Proteobacteria Gammaproteobacteria Proteobacteria Gammaproteobacteria Deltaproteobacteria Deltaproteobacteria 0.52 Alphaproteobacteria Alphaproteobacteria Unclassified Unclassified Unclassified Unclassified Verrucomicrobia Verrucomicrobiae Verrucomicrobia Verrucomicrobiae Low abundance 0.50 Low abundance

0.50 0.8 0.6 0.4 0.2 0.0 0.2 0.4 0.6 0.8 0.8 0.6 0.4 0.2 0.0 0.2 0.4 0.6 0.8 Proportion Proportion (a) (b) ImProm-II 42°C : TGIRT TGIRT : SuperScriptIV Phylum Class Phylum Class 0.64 Acidobacteria Acidobacteriia Acidobacteria Acidobacteriia Blastocatellia Blastocatellia Holophagae Holophagae Subgroup Subgroup 0.62 Thermoanaerobaculia Thermoanaerobaculia 0.62 Actinobacteria MB-A2-108 Actinobacteria MB-A2-108 Rubrobacteria Rubrobacteria Nitriliruptoria Nitriliruptoria 0319-7L14 0319-7L14 Thermoleophilia Thermoleophilia Acidimicrobiia 0.60 Acidimicrobiia Actinobacteria Actinobacteria 0.60 Armatimonadetes uncultured Armatimonadetes uncultured Fimbriimonadia Fimbriimonadia Bacteroidetes Bacteroidia Bacteroidetes Bacteroidia Chloroflexi Dehalococcoidia Chloroflexi Dehalococcoidia Ktedonobacteria 0.58 Ktedonobacteria TK10 TK10 0.58 KD4-96 KD4-96 JG30-KF-CM66 JG30-KF-CM66 Gitt-GS-136 Gitt-GS-136 Anaerolineae Anaerolineae Chloroflexia Chloroflexia Cyanobacteria Oxyphotobacteria 0.56 Cyanobacteria Oxyphotobacteria Deinococcus-Thermus Deinococci Deinococcus-Thermus Deinococci 0.56 Entotheonellaeota Entotheonellia Entotheonellaeota Entotheonellia Fibrobacteres Fibrobacteria Fibrobacteres Fibrobacteria Firmicutes Bacilli Firmicutes Bacilli Nitrospira Nitrospira Gemmatimonadetes S0134 0.54 Gemmatimonadetes S0134 BD2-11 BD2-11 0.54 Gemmatimonadetes Gemmatimonadetes Longimicrobia Longimicrobia Patescibacteria Saccharimonadia Patescibacteria Saccharimonadia Phycisphaerae Phycisphaerae Planctomycetacia 0.52 Planctomycetacia Proteobacteria Gammaproteobacteria Proteobacteria Gammaproteobacteria 0.52 Deltaproteobacteria Deltaproteobacteria Alphaproteobacteria Alphaproteobacteria Unclassified Unclassified Unclassified Unclassified Verrucomicrobia Verrucomicrobiae Verrucomicrobia Verrucomicrobiae Low abundance 0.50 Low abundance 0.50

0.8 0.6 0.4 0.2 0.0 0.2 0.4 0.6 0.8 0.8 0.6 0.4 0.2 0.0 0.2 0.4 0.6 0.8 Proportion Proportion (c) (d)

Figure S2. A proportional comparison of most abundant classes between the ImProm-II at 42 ◦C and SuperScriptIV (a),the ImProm-II at 55 ◦C and TGIRT, (b), ImProm-II at 42 ◦C and TIGRT (c) and TGIRT and SuperScriptIV (d). Enrichment is expressed such that a class that is equally proportional in both conditions, has a value of 0. If the class shows in one condition, but its absent from another, its value would be equal to 1 or -1 respectively. Furthermore a weighted average of the GC content of each class is expressed as the bar color. Each value is an average of 4 biological replicates.

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