Oligonucleotide primers used for gene expression analyses by RT-PCR
| Gene | 5′ Primer sequence | 3′ Primer sequence | Product size (ab) | Cycles1-a |
|---|---|---|---|---|
| Adseverin/Ads D5 | 5′-TGGCTTTGGGAAGGTGTACATCA | 5′-TGAGATGGGAAACAGCAAGGCAT | 1595/1296 | 25 |
| Adseverin | 5′-TGGCTTTGGGAAGGTGTACATCA | 5′-GCCATTGTTTCGTGGCAGTTTTA | 591 | 25 |
| CYP1A1 | 5′-GGCATTCATCCTTCGTCCCCTT | 5′-TCACAGCGGGCGTGTTTTAAAGT | 384 | 30 |
| Gelsolin | 5′-AAGGCAAGCAGGCCAACATGGA | 5′-CAAAGAGTCGAGGAGGATGGGCAT | 977 | 25 |
| IL-9 | 5′-CATCCTTGCCTCTGTTTTGCTCTTCA | 5′-TCTTCATGGTCGGCTTTTCTGCC | 427 | 30 |
| HPRT1-b | 5′-CACAGGACTAGAACACCTGC | 5′-GCTGGTGAAAAGGACTCT | 249 | 25 |
↵1-a The number of cycles used in the PCR reactions was optimized to ensure that analyzes of gene expression levels were done in the exponential phase.
↵1-b To normalize the amount of cDNA used in the PCR reactions, the housekeeping gene HPRT was amplified in the same reaction as the target gene as an internal control. For CYP1A1 and IL-9, the HPRT primers were included during the last 25 PCR cycles.