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Table 2 Polymerase chain reaction primer and probe sequences, amplicon sizes, and efficiencies

From: Accelerated cellular senescence in degenerate intervertebral discs: a possible role in the pathogenesis of intervertebral disc degeneration

Standard polymerase chain reaction conditions
Target Forward primer Reverse primer Amplicon size
GAPDH 5' CCC ATC ACC ATC TTC CAG G 3' 5' GGC CAT CCA CAG TCT TCT G 3' 354 bp
hTERT 5' GCC TGA GCT GTA CTT TGT CAA 3' 5' AGG CTG CAG AGC AGC GTG GAG AGG 3' 422 bp
Real-time polymerase chain reaction primers and probes
Target Forward primer Probe Reverse primer Efficiency
18s PDAR PDAR PDAR 99.65%
P16INK4a 5' GGC TCT ACA CAA GCT TCC TTT CC 3' 5' 6 FAM – CCC CCA CCC TGG CTC TGA CCA – TAMRA 5' TCA TGA CCT GCC AGA GAG AAC A 3' 99.22%
MMP-13 5' CCC CAG GCA TCA CCA TTC AAG 3' 5' 6 FAM – AGG GGT CCT GGC TGC CTT CCT CTT C – TAMRA 3' 5' GAC AAA TCA TCT TCA TCA CCA CCA C 3' 99.77%
ADAMTS 5 5' GGA CCT ACC ACG AAA GCA GAT C 3' 5' 6 FAM – CCC AGG ACA GAC CTA CGA TGC CAC C – TAMRA 3' 5' GCC GGG ACA CAC GGA GTA 3' 99.74%
  1. ADAMTS 5, a disintegrin and metalloprotease with thrombospondin motifs 5; bp, base pairs; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; hTERT, human telomerase reverse transcriptase; MMP-13, matrix metalloproteinase-13; PDAR, pre-developed assay reagent.