Table 2 Real-Time qPCR and Methylation-Specific PCR primers
Real-Time qPCR primers | ||||
Gene | NCBI ref seq | Sequence 5'-- > 3' | Ta, °C | Amplicon, bp |
Actb | NM_001101 | fw CTGGACTTCGAGCAAGAGATG | 60 | 180 |
rv AGTTGAAGGTAGTTTCGTGGATG | ||||
Cd80 | NM_005191 | fw CTCACTTCTGTTCAGGTGTTATCCA | 62 | 121 |
rv TCCTTTTGCCAGTAGATGCGA | ||||
Dnmt1 | NM_001130823 | fw TACCTGGACGACCCTGACCTC | 60 | 103 |
rv CGTTGGCATCAAAGATGGACA | ||||
Dnmt3a | NM_175629 | fw GACAAGAATGCCACCAAAGC | 60 | 190 |
rv CGTCTCCGAACCACATGAC | ||||
Dnmt3b | NM_006892 | fw GGCAAGTTCTCCGAGGTCTCTG | 60 | 113 |
rv TGGTACATGGCTTTTCGATAGGA | ||||
Methylation specific PCR primers | ||||
Gene | sequence 5'-- > 3' | Ta,°C | Amplicon, bp | |
CDH13 meth | Fw TCGCGGGGTTCGTTTTTCGC | 66 | 243 | |
Rv GACGTTTTCATTCATACACGCG | ||||
CDH13 unmeth | Fw TTGTGGGGTTGTTTTTTGT | 55 | 242 | |
Rv AACTTTTCATTCATACACACA | ||||
APC meth | Fw TATTGCGGAGTGCGGGTC | 64 | 98 | |
Rv TCGACGAACTCCCGACGA | ||||
APC unmeth | Fw GTGTTTTATTGTGGAGTGTGGGTT | 62 | 108 | |
Rv CCAATCAACAAACTCCCAACAA | ||||
RUNX3 meth | Fw TTACGAGGGGCGGTCGTACGCGGG | 71 | 220 | |
Rv AAAACGACCGACGCGAACGCCTCC | ||||
RUNX3 unmeth | Fw TTATGAGGGGTGGTTGTATGTGGG | 64 | 220 | |
Rv AAAACAACCAACACAAACACCTCC | ||||
MGMT meth | Fw TTTCGACGTTCGTAGGTTTTCGC | 64 | 81 | |
Rv GCACTCTTCCGAAAACGAAACG | ||||
MGMT unmeth | Fw TTTGTGTTTTGATGTTTGTAGGTTTTTGT | 64 | 93 | |
Rv AACTCCACACTCTTCCAAAAACAAAACA | ||||
MLH1 meth | Fw ACGTAGACGTTTTATTAGGGTCGC | 56 | 115 | |
Rv CCTCATCGTAACTACCCGCG | ||||
MLH1 unmeth | Fw TTTTGATGTAGATGTTTTATTAGGGTTGT | 56 | 124 | |
Rv ACCACCTCATCATAACTACCCACA | ||||