Table 1 Primer sequences of target genes using for RT-qPCR
Gene | Accession number | Sequences | (5’-3’) | Length (bp) | Tm (°C) |
|---|---|---|---|---|---|
Stemness genes | |||||
zinc finger protein 42 (ZFP42 or REX1) | XM_003639567.1 | Forward Reverse | AGGTTCTCACAGCAAGCTCA CCAGCAAATTCTGCGCACTG | 199 | 59.24 60.73 |
octamer-binding transcription factor 4 (OCT4) | XM_538830.1 | Forward Reverse | AGGAGAAGCTGGAGCAAAACC GTGATCCTCTTCTGCTTCAGGA | 100 | 60.55 59.50 |
Proliferation marker | |||||
proliferation marker protein Ki-67 (KI67) | XM_014108788.1 | Forward Reverse | GTGCAACTAAAGCACGGAGA GAGATTCCTGTTTGCGTTTTCGT | 124 | 58.49 58.49 |
Osteogenic markers | |||||
runt-related transcription factor 2 (RUNX2) | XM_005642335.1 | Forward Reverse | GGAAGAGGCAAGAGTTTCACC GTGCTCACTTGCCAACAGAA | 209 | 58.84 58.89 |
osterix (OSX) | XM_844688.3 | Forward Reverse | GCGTCCTCCCTGCTTGAG GCTTTGCCCAGTGTCGTTG | 122 | 60.13 60.01 |
collagen type I alpha 1 chain (COL1A1) | NM_001003090.1 | Forward Reverse | CCAGCCGCAAAGAGTCTACAT CTGTACGCAGGTGACTGGTG | 150 | 60.41 60.67 |
osteocalcin (OCN) | XM_547536.4 | Forward Reverse | GCCAGCCTATGGTCTCCTCTG CCACCAGCTCCTTCTGTTCTCT | 249 | 61.90 54.55 |
osteopontin (OPN) | XM_003434024.2 | Forward Reverse | GCCACAGAGCAAGGAAAACTC CTGCTTCTGAGATGGGTCAGG | 180 | 59.73 60.13 |
Reference gene | |||||
glyceraldehyde 3-phosphate dehydrogenase (GAPDH) | NM_001003142.1 | Forward Reverse | CCAACTGCTTGGCTCCTCTA GTCTTCTGGGTGGCAGTGAT | 100 | 59.38 59.67 |