Table 1.
Nucleotide sequences, amplification sites, GenBank accession numbers, amplimer sizes and PCR reaction conditions for each primer pair are shown.
| Gene | Primer | Nucleotide Sequence (5′→3′) | GenBank ID | Annealing Temperature | Amplimer Size (bp) |
|---|---|---|---|---|---|
|
Alkaline phosphatase
(Alpl) |
sense | GAA GTC CGT GGG CAT CGT (474–491) |
NM013059 | 59 °C | 347 |
| antisense | CAG TGC GGT TCC AGA CAT AG (801–820) |
||||
|
BMP2
(Bmp2) |
sense | AAG CCA GGT GTC TCC AAG (697–714) |
NM017178.1 | 53 °C | 209 |
| antisense | AAG TCC ACA TAC AAA GGG TG (886–905) |
||||
|
BMP4
(Bmp4) |
sense | TAG TCC CAA GCA TCA CCC (876–893) |
NM012827.2 | 53 °C | 294 |
| antisense | TCG TAC TCG TCC AGA TAC AAC (1149–1169) |
||||
|
BMP6
(Bmp6) |
sense | CCC AGA TTC CTG AGG GTG A (936–954) |
NM013107.1 | 56 °C | 248 |
| antisense | CAT GTT GTG CTG CGG TGT (1166–1183) |
||||
|
BMP7
(Bmp7) |
sense | AGG GAG TCC GAC CTC TTC T (607–625) |
NM001191856.1 | 54 °C | 297 |
| antisense | GTT CTG GCT GCG TTG TTT (886–903) |
||||
|
BMPR1
(Bmpr1a) |
sense | CCA TTG CTT TGC CAT TAT (240–257) |
NM009758.4 | 47 °C | 487 |
| antisense | TTT ACC AAC CTG CCG AAC (709–726) |
||||
|
Collagen type I
(Col1a1) |
sense | GGG CGA GTG CTG TGC TTT (348–365) |
NM007742.3 | 60 °C | 388 |
| antisense | GGG ACC CAT TGG ACC TGA A (717–735) |
||||
|
CREB
(Creb1) |
sense | AGA TTG CCA CAT TAG CCC (95–112) |
NM031017.1 | 52 °C | 441 |
| antisense | GCT GTA TTG CTC CTC CCT (518–535) |
||||
|
Actin
(Actb) |
sense | GCC AAC CGT GAA AAG ATG A (419–437) |
NM007393.5 | 54 °C | 462 |
| antisense | CAA GAA GGA AGG CTG GAA AA (861–880) |
||||
|
Gli1
(Gli1) |
sense | CCA CCC TAC CTC TGT CTA TTC G (2201–2222) |
NM010296.2 | 49 °C | 423 |
| antisense | CAC CCT TGT TCT GGT TTT ACC (2603–2623) |
||||
|
IHH
(Ihh) |
sense | CCA ACT ACA ATC CCG ACA TCA (248–268) |
NM053384.1 | 58 °C | 477 |
| antisense | GTC TTC ATC CCA GCC TTC C (390–408) |
||||
|
Osterix
(Sp7) |
sense | GCC TAC TTA CCC GTC TGA CTT T (525–543) |
NM001037632.1 | 56 °C | 131 |
| antisense | GCC CAC TAT TGC CAA CTG C (634–652) |
||||
|
Osteocalcin
(Bglap2) |
sense | TAA GGT GGT GAA TAG ACT CCG (123–143) |
NM013414.1 | 56 °C | 143 |
| antisense | CCT GGA AGC CAA TGT GGT (248–265) |
||||
|
PAC1
(ADCYAP1R1) |
sense | CTA CGC CCT TTA CTA CCC AG (210–229) |
NM016989.2 | 49 °C | 247 |
| antisense | GTA TTT CTT GAC AGC CAT TTG T (435–456) |
||||
|
PKA
(Prkaca) |
sense | GCA AAG GCT ACA ACA AGG C (847–865) |
NM008854 | 53 °C | 280 |
| antisense | ATG GCA ATC CAG TCA ATC G (1109–1126) |
||||
|
Osteopontin
(Spp1) |
sense | GCT GAA GCC TGA CCC ATC T (126–144) |
X51834 | 59 °C | 494 |
| antisense | TCC CGT TGC TGT CCT GAT (602–619) |
||||
|
PTCH1
(Ptch1) |
sense | GGA ACT TAT CAC GGA GAC AG (579–578) |
NM053566.1 | 56 °C | 368 |
| antisense | AAC CTT GAC ATC CAC CAT T (928–946) |
||||
|
Runx2
(Runx2) |
sense | GGA CGA GGC AAG AGT TTC A (598–616) |
NM001278483.1 | 55 °C | 249 |
| antisense | TGG TGC AGA GTT CAG GGA G (828–846) |
||||
|
SHH
(Shh) |
sense | TCG TGC TAC GCA GTC ATC G (1042–1060) |
NM017221.1 | 56 °C | 156 |
| antisense | CCT CGC TTC CGC TAC AGA (1180–1197) |
||||
|
Smad1
(Smad1) |
sense | AGC ACC TAC CCT CAC TCC C (935–953) |
NM013130.2 | 56 °C | 306 |
| antisense | GAA ACC ATC CAC CAA CAC G (1222–1240) |
||||
|
VPAC1
(VIPR1) |
sense | GTT CTA TGG CAC GGT CAA (376–393) |
NM001097523 | 52 °C | 216 |
| antisense | AGC AAT GTT CGG GTT CTC (573–590) |
||||
|
VPAC2
(VIPR2) |
sense | TCG GAA CTA CAT CCA TCT (477–497) |
NM001014970 | 48 °C | 177 |
| antisense | TTT GCC ATA ACA CCA TAC (636–653) |