Table 2.
Quantitative PCR primer/probe sets used for identification of mRNA transcripts in laser capture microdissected VMN and arcuate nucleus samples in diestrous rats and day 14 pregnant rats
| Primer set | Sequence (5′–3′) | Amplicon | Accession number | Efficiencies |
|---|---|---|---|---|
| Pacap | Forward GGAAACCCGCTGCAAGACTTCTAC | 150 bp | M63006 | 1.02 |
| Reverse CAAGACTTTGCGGTAGGCTTCGTT | F 136-159 | |||
| Probe ACGCCTACGCCCTTTACTACCCAGCCGACA | R 285-262 | |||
| P 206-225 | ||||
| Glucokinase | Forward CGAGCTTCAAGGAGCGGTTTCAC | 81 bp | NM_012565 | 1.02 |
| Reverse CCTCCTCTGATTCGATGAAGGTGATTTCG | F: 1341-1363 | |||
| Probe AGTGTGCGCAGGCTGACACCCAACT | R: 1421-1393 | |||
| P: 1367-1391 | ||||
| Bdnf | Forward AGAAGGTTCGGCCCAACGAAGAAA | 103 bp | NM_012513 | 0.94 |
| Reverse AGAAAGAGCAGAGGAGGCTCCAA | F: 890-913 | |||
| Probe TCCCGGGTGATGCTCAGCAGTCAAGTGCCT | R: 992-969 | |||
| P: 940-968 | ||||
| β-actin | Forward AGATGACCCAGATCATGTTTGAGA | 86 bp | NM_031144 | 0.95 |
| Reverse ACCAGAGGCATACAGGGACAA | F: 434-457 | |||
| Probe TCAACACCCCAGCCATGTACGTAGCC | R: 519-499 | |||
| P: 461-486 |
PCR, polymerase chain reaction; VMN, ventromedial nucleus of the hypothalamus; Pacap, pituitary adenylate cyclase–activated polypeptide; Bdnf, brain-derived neurotrophic factor.