Table 2.

Oligonucleotide primers used for methylation‐specific polymerase chain reaction for the promoter regions of hPER1, hPER2, hPER3, hCRY1 and hCRY2

Gene	Location	Annealing temperature (°C)
PER1
PER2‐M (amplicon size, 298 bp; location, −1879 to −1582)
Forward: 5′‐ATT TAG GTT TAC GTG CGT TC‐3′	−1879 to −1860	56
Reverse: 5′‐CGA CTC AAA AAC GAA AAT CG‐3′	−1582 to −1601
PER1‐U (amplicon size, 318 bp; location, −1889 to −1572)
Forward: 5′‐TAG TAT TAG TAT TTA GGT TTA TGT GTG TTT‐3′	−1889 to −1860	56
Reverse: 5′‐AAC AAC AAT CCA ACT CAA AAA CAA AAA TCA‐3′	−1572 to −1601
PER2
PER2‐M (amplicon size, 140 bp; location, −178 to −39)
Forward: 5′‐GCG GTT TCG TTG CGG TTT AT‐3′	−178 to −159	62
Reverse: 5′‐GCC GAC GCC GTT TCA AAC CG‐3′	−39 to −58
PER2‐U (amplicon size, 160 bp; location, −188 to −29)
Forward: 5′‐GTG GTG TGG TGT GGT TTT GTT GTG GTT TAT‐3′	−188 to −159	58
Reverse: 5′‐ACA CCC CCA CAC CAA CAC CAT TTC AAA CCA‐3′	−29 to −58
PER3
PER3‐M (amplicon size, 182 bp; location, −281 to −453)
Forward: 5′‐CGG GAG TTT TGG GTA TTC GC‐3′	−453 to −444	56
Reverse: 5′‐CGA CCC GAC TAA CTA AAA CG‐3′	−281 to −301
PER3‐U (amplicon size, 207, location, −269 to −465)
Forward: 5′‐TGG GTG GTT GGG TGG GAG TTT TGG GTA TTT GT‐3′	−465 to −444	56
Reverse: 5′‐AAT CCA ACA CCA ACA ACC CAA CTA ACT AAA ACA‐3′	−269 to −301
CRY1
CRY1‐M (amplicon size, 166 bp; location, −420 to −255)
Forward: 5′‐TCG TTT TGT TTT TAG GGG TC‐3′	−420 to −401	56
Reverse: 5′‐GCA ACC GCC TAA AAA CGA CG‐3′	−255 to −274
CRY1‐U (amplicon size, 187 bp; location, −432 to −246)
Forward: 5′‐ATT TTG GGT GGG TTG TTT TGT TTT TAG GGG TT‐3′	−432 to −401	65
Reverse: 5′‐ACA CCC ACC ACA ACC ACC TAA AAA CAA CA‐3′	−246 to −274
CRY2
CRY2‐M (amplicon size, 150 bp; location, −308 to −159)
Forward: 5′‐GTT TAT TTT CGG TAT TTC GC‐3′	−308 to −289	55
Reverse: 5′‐TAA CGA TTA ACC CAA AAA CG‐3′	−159 to −178
CRY2‐U (amplicon size, 171 bp; location, −318 to −148)
Forward: 5′‐TTG AGA TTT GGT TTA TTT TTG GTA TTT TGT‐3′	−318 to −289	56
Reverse: 5′‐TAC CTT CCA CCT AAC AAT TAA CCC AAA AAC A‐3′	−149 to −178
BMAL1
BMAL1‐M (amplicon size, 138 bp; location, 30–167)
Forward: 5′‐GGA GGT GTT TGT TTA TTC GC‐3′	30 to 39	58
Reverse: 5′‐AAA TAC GCG AAA TCG CGT CG‐3′	153 to 167
BMAL1‐U (amplicon size, 161 bp; location, 18–178)
Forward: 5′‐TAG GTT AGG GAT GGA GGT GTT TGT TTA TTT GT‐3′	18 to 39	60
Reverse: 5′‐AAC CCC CAA CAA AAT ACA CAA AAT CAC ATC A‐3′	153 to 178

The locations indicated are base numbers corresponding to the transcriptional start sites of the circadian genes.