Primer sequences for PCR amplification of TINF2
| Fragment . | Exon . | Forward . | Reverse . | Product size, bp . |
|---|---|---|---|---|
| 1 | 1 | CCTCTTACCGCCCTTTTCC | CTTGTCAGGTGCTCGCATC | 600 |
| 2 | 2 + 3 | AGACTAGTTGGAAAAGGTCAGC | GGGCGACAGAGCAAGATTC | 463 |
| 3 | 4 + 5 | CAAATGGCCAGGATTACAGG | TTCTTATGCCCGGAGCCC | 455 |
| 4 | 6 | GGCTCCGGGCATAAGAAAC | TGAGGTGAGAGCAAG CAAAG | 617 |
| 5 | CAGAGCAAAAGG AGTGAGTG | TACAGTCACAGGAAGAAACAG | 611 | |
| 6 | GGAATCTCTGGAAAACTATCAG | ACACCCAGATAATCTGGCAG | 552 |
| Fragment . | Exon . | Forward . | Reverse . | Product size, bp . |
|---|---|---|---|---|
| 1 | 1 | CCTCTTACCGCCCTTTTCC | CTTGTCAGGTGCTCGCATC | 600 |
| 2 | 2 + 3 | AGACTAGTTGGAAAAGGTCAGC | GGGCGACAGAGCAAGATTC | 463 |
| 3 | 4 + 5 | CAAATGGCCAGGATTACAGG | TTCTTATGCCCGGAGCCC | 455 |
| 4 | 6 | GGCTCCGGGCATAAGAAAC | TGAGGTGAGAGCAAG CAAAG | 617 |
| 5 | CAGAGCAAAAGG AGTGAGTG | TACAGTCACAGGAAGAAACAG | 611 | |
| 6 | GGAATCTCTGGAAAACTATCAG | ACACCCAGATAATCTGGCAG | 552 |
Primers are designed to genomic accession number AL096870. Fragment 1 includes the 5′ untranslated region (UTR). Fragments 4, 5, and 6 overlap and cover the alternative splice regions detailed in the 3′ UTR and described as 2 different isoforms (isoform 1, nm_001099274; isoform 2, nm_012461). All PCR amplifications were performed using 2 mM magnesium with an annealing temperature of 58°C for 30 cycles. Exceptions to this were for fragments 2 and 3, where 10% DMSO was used and the annealing temperature was increased to 60°C.