Table 2.
Protocols for reverse transcription semi-quantitative PCR
| Gapdh |
Ctnnb1
(β-catenin) |
Lcn2 | Lcn2-R (Slc22a17/24p3-R) | hLCN2-R isoform B | |
|---|---|---|---|---|---|
| Accession number | NM_001289726.1 | NM_007614.3 | NM_008491.1 | NM_021551.4 | (NM_016609.4) |
| Forward primer (5‘→3‘) | AGGGCTCATGACCACAGT | TTAAACTCCTGCACCCACCAT | CCACCACGGACTACAACCAG | CAGCCACCTCCTAACCGCTGTG | GCCATTCGCCACTGCTAC |
| Reverse primer (5‘→3‘) | TGCAGGGATGATGTTCTG | AGGGCAAGGTTTCGAATCAA | AGCTCCTTGGTTCTTCCATACA | CTCCCACTAGGCTCAAAGGCTGCT | GGAGAAGAGCCCAAGGACAGA |
| Activation | 5 min 95oC | 5 min 95oC | 5 min 95oC | 5 min 95oC | 5 min 95oC |
| Cycle number | 22 | 27 | 30-32 | 27-28 | 22 |
| Denaturation | 30 sec 94oC | 30 sec 94oC | 30 sec 94oC | 30 sec 94oC | 30 sec 94°C |
| Annealing | 30 sec 60oC | 30 sec 60oC | 30 sec 60oC | 30 sec 60oC | 30 sec 55°C |
| Extension | 30 sec 72oC | 30 sec 72oC | 30 sec 72oC | 30 sec 72oC | 30 sec 72°C |
| Final Extension | 7 min 72oC | 7 min 72oC | 7 min 72oC | 7 min 72oC | 10 min 72°C |
| PCR product (bp) | 112 | 73 | 100 | 86 | 255 |
bp = base pairs