Table 1.

Gene PCR and pyrosequencing primers and PCR annealing temperature

GeneForward primerReverse primerPyrosequencing primerAnnealing temperature
CCNA1Biotin-TTTGCGTAGTTTCGAGGATTTCCCGTTCTCCCAACAACCGCTAACAACCCCCTCTA60°C
CYGBBiotin-TCGATCGTTAGTTCGTTCACTAACTCGAAAACGCGACCCAACTAAATCCAC56°C
P16CGGAGGGGGTTTTTTCGTTAGTATCBiotin-TCCCCCTCTCCGCAACCGGGTTGGTTATTAGAGGGT62°C
MGMTCGGTTTCGTTTTCGCGTTTCBiotin-ACCGCGAAAACCTACGAACGGGATATGTTGGGATAGT62°C
  • NOTE: Biotin: the indicated primers were 5′ biotin labeled to enable preparation of the appropriate ssDNA for subsequent pyrosequencing reactions.

    Abbreviations: CCNA1, cyclin A1; CYGB, cytoglobin; MGMT, 6-O-methylguanine DNA methyltransferase.