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Table 3 Reference genes, qPCR primers and reaction parameters.

From: Selection and evaluation of reference genes for improved interrogation of microbial transcriptomes: case study with the extremophile Acidithiobacillus ferrooxidans

Gene Forward primer (5'-3') Reverse primer (5'-3') Amplicon size Q-PCR Linear correlation coefficient Q-PCR Efficiency (%) Average Ct ± SD
Candidate reference genes
   coaE ACT ATC GCC CAT TGC TGG AT TTG GTA ATG ATC CAG TCG GC 169 0.999 93 25.50 ± 0.69
   era CAT GGA TGA GAT CAA GAG CG GTA TCC CGA GAA TCT GAT CC 174 0.999 94 22.56 ± 0.21
   gmk ATG GCA CTA GTG AAC CTT GG AAT GAC CGT CTC CGA ATC CT 197 0.998 92 24.49 ± 0.50
   gyrA TAC CTC GAT TAC GCC ATG AG TGT CAT AGA CAG CGG TAT CG 202 0.998 95 26.24 ± 0.45
   map TTA CCA CCG ATG AAC TGG AC AGC CAT CCT TGA TAA CCG TG 222 0.999 94 20.64 ± 0.29
   nth ATC GTC TGG GAC TGT TCA AC TAT TGA GGA CGA CAT TGG CG 153 0.997 94 25.31 ± 0.40
   rplI GAT GCC ATG GTT ACG ATT GC GAC CAT CAC ATC CAG TTC GA 210 1.000 92 21.68 ± 0.51
   rpoC AAT GCG GTG TTG AGG TAA CC AGG TAC TGG TCT TCG GTA AG 238 0.997 95 20.79 ± 0.23
   trpS TCT GCT CAT CGA ATG GTT GG GAA TGT CTG CCG TCA TCA AC 230 0.999 89 23.86 ± 0.70
Reference genes used in prior studies
   alaS CTG GAA TCT GGT CTT CAT GC GCT TGA AGA GAT CGG TGT CA 152 1.000 94 22.35 ± 0.39
   recA CCG CCA ACA TTT CCC GGA CC ACG CCG CGG TCC ACC AGT TC 301 0.998 98 29.41 ± 0.88
   rrs ACA CTG GGA CTG AGA CAC GG ACC GCC TAC GCA CCC TTT AC 277 0.999 95 8.93 ± 0.27
Differentially expressed target genes
   cbbOIa ACG GCA TTG AGC TAT ACC G TCC GGG TCT AGT AGT GCA T 265 0.996 97 21.55 ± 0.72
   cyoB CAA TTA CAT GGT CCA CAA CA AGC GTA TAC ACC ACG ATA CC 300 0.995 92 28.11 ± 2.17
   mntH ATA TCG GTG CCG TCA TCA TG GAG AGC TGA TGC ACC GTA TT 217 0.996 97 21.92 ± 2.02
   sdrAI GTT TGG GTG CTG AAG TAG TG GCA ACA GTG GCA AAC AGG C 255 0.995 93 23.61 ± 2.82
  1. Primer sequences and reaction parameters used for quantitative real time PCR expression analysis.