Table 1.
Sequences of hybridization probes and qRT-PCR primers
| Name | DNA Sequence |
|---|---|
| Hybridization Probes | |
| 5' 3' | |
| SCD1 (AS)1 | 1007 GTGGTGAAGTTGATGTGCCAGCGGTACTCACTG 975 |
| EIF3H (AS)2 | 1034 GGCAGTGAACTCCTTGATGTTCTGGCAGTAAGTGTT 999 |
| qRT-PCR Primers | |
| 5' 3' | |
| SCD1 (S)1 | 26 GAAGCGAGCAACCGACAGCCAC 47 |
| SCD1 (AS)1 | 180 GTCTTCTTCCAGATAGAGGGGCAC 157 |
| EIF3H (S)2 | 850 AACACCAGTATCAGCAGCGTCG 871 |
| EIF3H (AS)2 | 1027 AACTCCTTGATGTTCTGGCAGTAAGTG 1001 |
1 Sequence and numbering based on rat SCD1 (GenBank ID: NM_139192.2)
2 Sequence and numbering based on rat EIF3H (GenBank ID: NM_198751.1)
SCD1 and EIF3H hybridization probes are located within the protein-coding regions. The PCR-amplified sequence from EIF3H mRNA includes most of the 33-mer used as the EIF3H hybridization probe. The PCR-amplified sequence from SCD1 mRNA does not overlap with the SCD1 hybridization probe, because of the necessity to avoid potential cross reactivity with SCD2 mRNA, but it does produce an amplicon that is mostly within the protein-coding region. The SCD1 primer set does not match the SCD2 mRNA sequence (GenBank ID: NM_031841.1), and cloning and sequencing of the product generated by qRT-PCR confirmed that the amplified sequence was SCD1.