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Table 4 Primers used in quantitative reverse transcription - polymerase chain reaction (QPCR)

From: Heat-shock responsive genes identified and validated in Atlantic cod (Gadus morhua) liver, head kidney and skeletal muscle using genomic techniques

Primer name

Sequence

Gene name of the best BLASTx hit

Amplification efficiency

Amplicon size (bp)

HSP90α Forward

5'- GAA CAA GAC CAA GCC CCT TT -3'

Heat shock protein 90 alpha

107%

133

HSP90α Reverse

5'- CTG ACC CTC CAC CGA GAA GT -3'

   

GRP94 Forward

5'- AGT GTT TCT CTC GAC ACG TTC A -3'

Heat shock protein 90 kDa beta, member 1 (synonym: GRP94)

91%

97

GRP94 Reverse

5'- CAG ACG ACT TCC ATG ACA TGA T -3'

   

HSP70-1 Forward

5'- GAG AAC AAG ATC ACC ATC ACG A -3'

HSP70-1 (synonym: HSPA1A)

97%

123

HSP70-1 Reverse

5'- GGC TGT TAC TTT CTC TCC CTG A -3'

   

GRP78 Forward

5'- CTC CTT CAT TTT GGT CAG AAC C -3'

78 kDa glucose-regulated protein (synonym: GRP78)

90%

132

GRP78 Reverse

5'- CTC AAG TTC CTC CCA TTC AAA G -3'

   

HSP47 Forward

5'- ATG GAA GTC AGC CAC AAC CT -3'

Heat Shock Protein 47

93%

100

HSP47 Reverse

5'- TCT TGC CCG TGA TGT TAG AC -3'

   

CCT1 Forward

5'- GCA GGC GTT TGG GAT AAC TA -3'

T-complex-1

95%

127

CCT1 Reverse

5'- GCG CTT AAC CCT TCA GAG AA -3'

   

CCT5 Forward

5'- CCA GGC GAG GTT GAA GAA TA -3'

Chaperonin containing TCP1, subunit

92%

91

CCT5 Reverse

5'- TAG AAC AGG GGA GTG GTG GT -3'

5 (epsilon)

  

TCTP Forward

5'- ACC AAG CCA GAG AGA GTG GA -3'

Translationally-controlled tumor protein

101%

147

TCTP Reverse

5'- ATC CTC ACG GAA GTC AAG CA -3'

   

Enolase Forward

5'- GGA CGG CAC TGA AAA CAA AT -3'

Enolase 3

93%

115

Enolase Reverse

5'- ACA GAG GAA CCC CCT TCT CC -3'

   

PFK Forward1

5'- TGT TTG CCA ACT CCC CAG AGA -3'

Phosphofructokinase, muscle a

96%

121

PFK Reverse1

5'- TCC GGT GCT TGA AGT CTG TCA -3'

   

Aldolase Forward1

5'- TGA CAT TGC TCA GAG GAT GG -3'

Aldolase B

91%

143

Aldolase Reverse1

5'- TAG CGA CGG TTC TCC TCA CT -3'

   

Bikunin Forward

5'- GCC ACT GAG TTC ACA GAC G -3'

Alpha-1-microglobulin/bikunin precursor

90%

107

Bikunin Reverse

5'- CAG CTC ATG GAG GAG GAG T -3'

   

IgM Forward

5'- GAG CAT CCA CTG GCT CTT TA -3'

Immunoglobulin heavy chain, secretory form

93%

97

IgM Reverse

5'- GCA GCA AGC TAT ATC CAG GT -3'

   

Nupr1 Forward

5'- CTT TCT TCT CGC TGT TCT GC -3'

Nuclear protein 1

88%

101

Nupr1 Reverse

5'- GGA AGG ACC AAG AAG GAG TC -3'

   

IL-8 Forward

5'- CTT CAG CAT CCA GAC AGA CC -3'

Interleukin-8

100%

133

IL-8 Reverse

5'- CAG ACA GAG AGC CGT CAG AT -3'

   

TLR22 Forward

5'- TGC AGG TAA TCA CGA CTG AC -3'

TLR22

85%

93

TLR22 Reverse

5'- GAG ACT TCC AGC CAG ACC TA -3'

   

18S Forward

5'- ATG GCC GTT CTT AGT TGG TG -3'

18S ribosomal RNA (normalizer gene)

109%

180

18S Reverse

5'- GGA CAT TTA AGG GCG TCT CA -3'

   
  1. 1The primers for aldolase and PFK were designed to amplify a region that is conserved between the muscle and liver forms of these genes.