Table 2 Primers and polymerase chain reaction (PCR) conditions for quantitative reverse transcription PCR
Region/gene | Primer, forward | Primer, reverse | PCR conditions |
|---|---|---|---|
Xist, Amp 4 | TN4s ttctaccctttcctctcctcatc | JTL4as, gaggtacgtaagctcagtga | 95°C for 5 minutes; (95°C for 30 seconds; 60°C for 30 seconds; 72°C for 30 seconds) × 40 cycles |
Xist, Amp 5 | Qmex4, gcaaggaagacaaaggctcaaagaat | Qmex52 ggagagagaaccaaatagagcagaat | 95°C for 3 minutes; (95°C for 20 seconds; 61°C for 15 seconds; 72°C for 30 seconds) × 40 cycles |
Xist, Amp 51 | DE-SOL2, tgcaatctttgtggccactcctcttctg | TN51, tatcaaaacgtcaaaaatctcg | 95°C for 5 minutes; (95°C for 30 seconds; 60°C for 30 seconds; 72°C for 30 seconds) × 40 cycles |
Xist mut, Amp 51m | DE-SOL2, tgcaatctttgtggccactcctcttctg | neoTN9, catcgcattgtctgagtaggtgtc | 95°C for 5 minutes; (95°C for 30 seconds; 64°C for 30 seconds; 72°C for 30 seconds) × 40 cycles |
Dnmt1 | Dnmt1_F1, agatccactgtggcaagaaga | Dnmt1_R1, ctgaagttcaccacagcttcc | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt1 | Dnmt1_F2, agggaccatatctgcaaggac | Dnmt1_R2, gctgctgtagccatttttcac | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt3a2 | Dnmt3a2_F1, cagacgggcagctatttacag | Dnmt3a2_R1, ggttctcttccacagcattca | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt3a2 | Dnmt3a2_F2, ggctcacacctgagctgtact | Dnmt3a2_R2, cctcctccaccttctgagact | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt3b | Dnmt3b_F1, caagcgcctcaagacaaatag | Dnmt3B_R1, gcgatcccggcaactctgaca | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt3b | Dnmt3b_F2, cgagaacaaaagtcgaagacg | Dnmt3b_R2, gggttcttctttccacaggac | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt3b | Dnmt3b_F3, ccattcttctggatgttcgag | Dnmt3b_R3, tctgatggagttcgacttggt | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt3L | Dnmt3L_F1, cttgtttgagggagggttatg | Dnmt3L_R1, gtacagtcggggctctcacag | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt3L | Dnmt3L_F2, agacaactacccgcttccttc | Dnmt3L_R2, ctcttcttcctttggggtcag | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Dnmt3L | Dnmt3L_F3, cccctaggcagctcttgtgat | Dnmt3L_R3, gcgggtagttgtctcttggtc | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
Idh2 | Idh1F, agaaaatgtggaagagccctaacg | Idh1R, tgccagctcgatctaccacaaaat | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |
β-actin | BA11, gatatcgctgcgctggtcgt | BA2, agatcttctccatgtcgtcc | 95°C for 3 minutes; (95°C for 20 seconds; 60°C for 20 seconds) × 40 cycles |