Table 1.

Genes selected for methylation analysis in acute myeloid leukemia

GeneChromosomeFunctionPrimers (F/R)Annealing (temperature, no. cycles)Restriction enzyme
CACNA1G 17q21 T-type calcium channel GGGGGYGTTTTTTTTYGGATTTT 58 (5), 56 (5), 54 (5), 52 (26), 30 sec EcoRI 
TTCCCCTACRCCCCTAAAACTTCC  
ER-alpha 6q25 Estrogen receptor GGTTTTTGAGTTTTTTGTTTTG 60 (3), 57 (4), 54 (5), 51 (25), 30 sec TaqI  
AACTTACTACTATCCAAATACACCTC 
GPR37 7q31 G-protein–coupled receptor GGTTAGGTGGGGTAAGAGTTT 56, 45 sec, 35 cycles HinfI  
AACRTTTAATCCAATTACAAACC 
MDR1 7q21 Drug efflux GTTATAGGAAGTTTGAGTTT 54 (3), 51 (4), 48 (5), 45 (26), 30 sec Taq
AAAAACTATCCCATAATAAC  
MINT1 5q13 CpG island hypermethylated in cancer GGGTTGGAGAGTAGGGGAGTT 55 (35) 30 sec TaqI  
CCATCTAAAATTACCTCRATAACTTA 
MINT2 2p22 CpG island hypermethylated in cancer YGTTATGATTTTTTTGTTTAGTTAAT 60 (3), 58 (4), 56 (5), 54 (26) TaqI  
TACACCAACTACCCAACTACCTC 
MLH1 3p21 Mismatch repair TAGTAGTYGTTTTAGGGAGGGA 53 (35) 30 sec RsaI  
TCTAAATACTCAACRAAAATACCTT 
MyoD 11p15 Muscle-specific transcription factor TGTTGGAGAGGTTTGGAAAGG 53, 30 sec, 35 cycles RsaI  
AAAATCCRAAACCAATAAAAACAC 
p15 9p21 Cyclin-dependent kinase inhibitor GGAGTTTAAGGGGGTGGG 50, 30 sec, 35 cycles NruI  
CCTAAATTACTTCTAAAAAAAAAC 
p16 9p21 Cyclin-dependent kinase inhibitor GGTTTTGGYGAGGGTTGTTT 58 (5), 56 (5), 54 (5), 52 (26), 30 sec TaqI  
ACCCTATCCCTCAAATCCTCTAAAA 
PITX2 4q25 Homeotic gene TAAGTGTTTYGTTAGGTTTTTT 50, 30 sec, 35 cycles TaqI  
CCAAACTCCACTACACAATAAC 
PTC-A 9q22 WNT signaling GTTGGTTTGTTAATYGGAGT 58 (5), 56 (5), 54 (5), 52 (26), 30 sec TaqI  
TTACCAACCRAAACCATATT 
PTC-B 9q22 WNT signaling 5-AATGTGTGGAATTTAGGGA 54, 45 sec, 35 cycles HgaI  
TAAAACAACCAACTACAACTTAC 
SDC4 20q12 Cell surface heparan sulfate proteoglycan GGGGATTTGTTTGGTAGTGG 57, 45 sec, 35 cycles HinfI  
CCCGAAATTCCCCAATAAA 
THBS1 15q15 Angiogenesis inhibitor GGAGAGAGGAGTTTAGATTGGTT 54, 45 sec, 38 cycles BstUI  
AAATAAAAATTACTCCTAAAAAAC 
GeneChromosomeFunctionPrimers (F/R)Annealing (temperature, no. cycles)Restriction enzyme
CACNA1G 17q21 T-type calcium channel GGGGGYGTTTTTTTTYGGATTTT 58 (5), 56 (5), 54 (5), 52 (26), 30 sec EcoRI 
TTCCCCTACRCCCCTAAAACTTCC  
ER-alpha 6q25 Estrogen receptor GGTTTTTGAGTTTTTTGTTTTG 60 (3), 57 (4), 54 (5), 51 (25), 30 sec TaqI  
AACTTACTACTATCCAAATACACCTC 
GPR37 7q31 G-protein–coupled receptor GGTTAGGTGGGGTAAGAGTTT 56, 45 sec, 35 cycles HinfI  
AACRTTTAATCCAATTACAAACC 
MDR1 7q21 Drug efflux GTTATAGGAAGTTTGAGTTT 54 (3), 51 (4), 48 (5), 45 (26), 30 sec Taq
AAAAACTATCCCATAATAAC  
MINT1 5q13 CpG island hypermethylated in cancer GGGTTGGAGAGTAGGGGAGTT 55 (35) 30 sec TaqI  
CCATCTAAAATTACCTCRATAACTTA 
MINT2 2p22 CpG island hypermethylated in cancer YGTTATGATTTTTTTGTTTAGTTAAT 60 (3), 58 (4), 56 (5), 54 (26) TaqI  
TACACCAACTACCCAACTACCTC 
MLH1 3p21 Mismatch repair TAGTAGTYGTTTTAGGGAGGGA 53 (35) 30 sec RsaI  
TCTAAATACTCAACRAAAATACCTT 
MyoD 11p15 Muscle-specific transcription factor TGTTGGAGAGGTTTGGAAAGG 53, 30 sec, 35 cycles RsaI  
AAAATCCRAAACCAATAAAAACAC 
p15 9p21 Cyclin-dependent kinase inhibitor GGAGTTTAAGGGGGTGGG 50, 30 sec, 35 cycles NruI  
CCTAAATTACTTCTAAAAAAAAAC 
p16 9p21 Cyclin-dependent kinase inhibitor GGTTTTGGYGAGGGTTGTTT 58 (5), 56 (5), 54 (5), 52 (26), 30 sec TaqI  
ACCCTATCCCTCAAATCCTCTAAAA 
PITX2 4q25 Homeotic gene TAAGTGTTTYGTTAGGTTTTTT 50, 30 sec, 35 cycles TaqI  
CCAAACTCCACTACACAATAAC 
PTC-A 9q22 WNT signaling GTTGGTTTGTTAATYGGAGT 58 (5), 56 (5), 54 (5), 52 (26), 30 sec TaqI  
TTACCAACCRAAACCATATT 
PTC-B 9q22 WNT signaling 5-AATGTGTGGAATTTAGGGA 54, 45 sec, 35 cycles HgaI  
TAAAACAACCAACTACAACTTAC 
SDC4 20q12 Cell surface heparan sulfate proteoglycan GGGGATTTGTTTGGTAGTGG 57, 45 sec, 35 cycles HinfI  
CCCGAAATTCCCCAATAAA 
THBS1 15q15 Angiogenesis inhibitor GGAGAGAGGAGTTTAGATTGGTT 54, 45 sec, 38 cycles BstUI  
AAATAAAAATTACTCCTAAAAAAC 
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