Table 1.

Primer sequences, annealing temperature, and product sizes for methylation-specific polymerase chain reaction assays

CpG status	Genes	Forward primer (5’→3’)	Reverse primer (5’→3’)	Genomic position2	Annealing temperature (°C)	Product size (bp)
M	hMLH1	ACGTAGACGTTTTATTAGGGTCGC	CCTCATCGTAACTACCCGCG	-716 to -602	55	115
U		TTTTGATGTAGATGTTTTATTAGGGTTGT	ACCACCTCATCATAACTACCCACA	-721 to -598	55	124
M	MGMT1	TTTCGACGTTCGTAGGTTTTCGC	GCACTCTTCCGAAAACGAAACG	+142 to +223	62	121
U		TTTGTGTTTTGATGTTTGTAGGTTTTTGT	AACTCCACACTCTTCCAAAAACAAAACA	+137 to +230	62	133
M	CDKN2A/p16	TTATTAGAGGGTGGGGCGGATCGC	CCACCTAAATCGACCTCCGACCG	+167 to +401	65	234
U		TTATTAGAGGGTGGGGTGGATTGT	CCACCTAAATCAACCTCCAACCA	+167 to +401	55	234

¹All of the primers for the MGMT gene were modified with a 20 bp GC-rich tail (5’-GCGGTCCCAAAAGGGTCAGT-3’) at their 5’ end;

²GenBank (PubMed). M: Methylated; U: Unmethylated; hMLH1: AB017806; MGMT: DD183709; CDKN2A/p16: DQ406745.