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Table 2 Nucleotide sequences for the specific primers and MGBâ„¢ probes, melting temperatures ( T m ), fragment lengths and PCR efficiencies

From: Convergent evolution of heat-inducibility during subfunctionalization of the Hsp70 gene family

Target

Primer/Probe

Nucleotide sequence

T m

Fragment length

PCR efficiency

Hsp70 CY-A

Hsp70cyA_rt_F

5′-TCA GGA GCT GAT GAC AAA CCA A-3′

59°C

74 bp

90%

 

Hsp70cyA_rt_R

5′-ATC TCC TCT GGA TGG AAT TTC TTG-3′

59°C

  
 

Hsp70cyA_MGB_VIC

5′-TGT GGT TAA GTA TAA AGG TGA G-3′

67°C

  

Hsp70 CY-B

Hsp70cyB_rt_F4

5′-TGC ATA TGG TGC TGC TGT TTA AG-3′

59°C

133 bp

89%

 

Hsp70cyB_rt_R2

5′-TAA TAC ACT CAT TAC ACC ACC TGC AG-3′

59°C

  
 

Hsp70cyB_MGB_VIC

5′-CCA CTT TCA TTA GGA ATT-3′

68°C

  

Hsp70 ER-A

Hsp70ERA_rt_F

5′-GAA GTT CTC ACT AGA GCC AGA TTC G-3′

59°C

76 bp

93%

 

Hsp70ERA_rt_R

5′-CTG ATT ACA TGG GAC CAG TTG TCT-3′

58°C

  
 

Hsp70ERA_MGB_VIC

5′-ACT CAA TTC AGA TCT C-3′

68°C

  

Hsp70 ER-B

Hsp70ERB_rt_F

5′-GTT GAG AAG GGA ACC TAA TAA AAG GTC-3′

59°C

98 bp

92%

 

Hsp70ERB_rt_R

5′-GTC CCA AAT ATC CTT CAG CAA TCT-3′

59°C

  
 

Hsp70ERB_MGB_VIC

5′-CAG AGG AGA TTA GTG CCA TG-3′

68°C

  

Hsp70 MT

Hsp70mt_rt_F

5′-ACC AGA ATG CCA AAA GTC CAA-3′

58°C

76 bp

91%

 

Hsp70mt_rt_R

5′-CAT CTG GGT TGA CTG ACT TGT TG-3′

58°C

  
 

Hsp70mt_MGB_VIC

5′-AAG ATT TAT TCG ACA AAC CA-3′

67°C

  

GAPDH

GAPDH_rt_F

5′-GCT GCC AAG GCT GTT GGT-3′

58°C

75 bp

92%

 

GAPDH_rt_R

5′-TGT TGG AAC TCT GAA GGC CAT AC-3′

59°C

  
 

GAPDH_MGB_FAM

5′-TCC CAG AAA TCA AAG-3′

67°C

  

EF-1a

EF1a_rtF2

5′-TTG ATG CCC CAG GAC ATA GAG-3′

59°C

95 bp

94%

 

EF1a_rtR2

5′-TCC TGC TGG TGA GGC AAT C-3′

60°C

  
 

EF1a_MGB_FAM

5′-TAC AGG AAC ATC ATA AGC-3′

68°C

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