Table 1.
Primers used to amplify cDNAs encoding mouse or human HSP27, mouse GAPDH, and proinflammatory cytokines based on published GenBank sequences for mice
| Primer | Accession number | Sequence (sense, antisense) | Product size (bp) | Cycle number | Annealing T (°C) |
|---|---|---|---|---|---|
| Mouse HSP27 | NM_024441 | 5′-CCTAAGGTCTGGCATGGTA-3′ | 373 | 26 | 66 |
| 5′-AGGAAGCTCGTTGTTGAAGC-3′ | |||||
| Human HSP27 | NM_001540 | 5′-GTCCCTGGATGTCAACCAC-3′ | 259 | 21 | 66 |
| 5′-GACTGGGATGGTGATCTCG-3′ | |||||
| Mouse KC | J04596 | 5′-CAATGAGCTGCGCTGTCAGTG-3′ | 203 | 23 | 60 |
| 5′-CTTGGGGACACCTTTTAGCATC-3′ | |||||
| Mouse MIP-2 | X53798 | 5′-CCAAGGGTTGACTTCAAGAAC-3′ | 282 | 22 | 60 |
| 5′-AGCGAGGCACATCAGGTACG-3′ | |||||
| Mouse ICAM-1 | X52264 | 5′-TGTTTCCTGCCTCTGAAGC-3′ | 409 | 21 | 60 |
| 5′-CTTCGTTTGTGATCCTCCG-3′ | |||||
| Mouse TNF-α | X02611 | 5′-CCTCAGCCTCTTCTCCTTCCT-3′ | 290 | 24 | 65 |
| 5′-GGTGTGGGTGAGGAGCA-3′ | |||||
| Mouse MCP-1 | NM_011333 | 5′-ACCACAGTCCATGCCATCAC-3′ | 312 | 22 | 60 |
| 5′-CACCACCCTGTTGCTGTAGCC-3′ | |||||
| Mouse GAPDH | M32599 | 5′-ACCACAGTCCATGCCATCAC-3′ | 450 | 15 | 65 |
| 5′-CACCACCCTGTTGCTGTAGCC-3′ |
ICAM-1, intercellular adhesion molecule-1; KC, keratinocyte-derived chemokine; MCP-1, monocyte chemoattractant protein 1; MIP-2, macrophage inflammatory protein 2; TNF-α, tumor necrosis factor-α.
Respective anticipated RT-PCR product size, PCR cycle number for linear amplification, and annealing temperatures used for each primer are also provided.