Table 1 PCR primers, cycling conditions and positive controls used.
Primer | Sens Primer | Antisense Primer | Size (bp)* | annealing (celsius) | cycles | extension | positive control |
|---|---|---|---|---|---|---|---|
Keratin 1 | 5'-gatgaaggccacggtgatca-3' | 5'-gacttgagttggggtgccta-3' | 627 | 64 | 36 | 1 min | KLE |
Keratin 4 | 5'-ctccagcaaaaaccttgagc-3' | 5'-aagtcattctcggctgctgt-3' | 186 | 58 | 36 | 30 sec | KLE |
Keratin 5 | 5'-tctcgccagtcaagtgtgtc-3' | 5'-atagccacccactccacaag-3' | 247 | 58 | 36 | 30 sec | KLE |
Keratin 7 | 5'-caggatgtggtggaggactt-3' | 5'-ttgctcatgtaggcagcatc-3' | 116 | 58 | 30 | 30 sec | KLE |
Keratin 8 | 5'-agatgaaccggaacatcagc-3' | 5'-tccagcagcttcctgtaggt-3' | 262 | 58 | 30 | 30 sec | KLE |
Keratin 13 | 5'-gtcttcagcacccagaggag-3' | 5'-ttgcagaaaggcaggaaact-3' | 246 | 58 | 36 | 30 sec | Hec-1-A |
Keratin 18 | 5'-cacagtctgctgaggttgga-3' | 5'-gagctgctccatctgtaggg-3' | 164 | 58 | 36 | 30 sec | KLE |
Keratin 19 | 5'-tttgagacggaacaggctct-3' | 5'-gccatgacctcatattggct-3' | 275 | 58 | 30 | 30 sec | KLE |
Keratin 20 | 5'-acgccagaacaacgaatacc-3' | 5'-acgaccttgccatccactac-3' | 198 | 61 | 36 | 30 sec | KLE |
Vimentin | 5'-gagaactttgccgttgaagc-3' | 5'-tccagcagcttcctgtaggt-3' | 170 | 58 | 36 | 30 sec | KLE |
Desmin | 5'-caagctgcaggaggagattc-3' | 5'-ggcagtgaggtctggcttag-3' | 241 | 62 | 36 | 30 sec | KLE |
ER alpha | 5'-gtgcctggctagagatcctg-3' | 5'-agagacttcagggtgctgga-3' | 265 | 66 | 36 | 30 sec | MCF-7 |
ER beta | 5'-tcaggcatgcgagtaacaag-3' | 5'-gcttttactgtcctctgccg-3' | 167 | 65 | 36 | 30 sec | MCF-7 |
PR-A | 5'-gcttcaagttagccaagaagagt-3' | 5'-ctggaaattcaacactcagtg-3' | 290 | 58 | 36 | 30 sec | Ishikawa |
PR-B | 5'-acaccttgcctgaagtttcg-3' | 5'-tccaagacactgtccagcag-3' | 160 | 64 | 35 | 30 sec | Ishikawa |
MMP-2 | 5'-aggcaagtggtccgtgtgaa-3' | 5'-acagtggacatggcggtctcag-3' | 369 | 66 | 36 | 30 sec | KLE |
MMP-9 | 5'-caacatcacctattggatcc-3' | 5'-cgggtgtagagtctctcgct-3' | 479 | 60 | 36 | 30 sec | KLE |
MMP-14 | 5'-ccagggtctcaaatggcaaca-3' | 5'-ccatggaagccctcggcaaa-3' | 219 | 66 | 36 | 30 sec | ** |
TIMP-1 | 5'-accagaccaccttataccagcg-3' | 5'-ggactggaagcccttttcagag-3' | 395 | 58 | 36 | 30 sec | RL-95-2 |
TIMP-2 | 5'-atgcagatgtagtgatcagggc-3' | 5'-gatgaagtcacagagggtgatg-3' | 272 | 58 | 36 | 30 sec | RL-95-2 |
β-actin | 5'-gaggatcttcatgaggtagtctgtcaggtc-3' | 5'-caactgggacgacatggagaagatctggca-3' | 348 | 58 | 25 | 30 sec | none |