TABLE 2.
Primers for nested Tlr amplificationa
| Receptor | Primer (sequence)
|
Product size (bp) | |
|---|---|---|---|
| Sense | Antisense | ||
| mMD2 | mMD2-S117 (5′-AGTGGTTCTGCAACTCCTCC) | mMD2-AS338 (5′-GGCACAGAACTTCCTTACGC) | 222 |
| mCD14 | mCD14-S317 (5′-GCGGATTCCTAGTCGGATTC) | mCD14-AS951 (5′-GGTTCCTATCCAGCCTGTTG) | 635 |
| mTLR-1 | mTLR1-S722 (5′-AATCTCTTCGGCACGTTAGC) | mTLR1-AS1049 (5′-ATCCTGAAGGCAAGTTGACC) | 328 |
| mTLR-2 | mTLR2-S658 (5′-TTATCTTCCTCCTGGTTCGG) | mTLR2-AS1339 (5′-TGATTCGCTTCACCTTCTCC) | 682 |
| mTLR-3 | mTLR3-S895 (5′-CAACAACCTCCATGATGTCG) | mTLR3-AS1412 (5′-TCAGACCTCTCCATTCCTGG) | 518 |
| mTLR-4 | mTLR4-S664 (5′-GACTTCATTCAAGACCAAGCC) | mTLR4-AS968 (5′-ACACCTGCCAGAGACATTGC) | 305 |
| mTLR-5 | mTLR5-S507 (5′-TTGTCCGATCATCAGCTAAGC) | mTLR5-AS867 (5′-GGCAGATTCTTCTTGTCTTGG) | 361 |
| mTLR-6 | mTLR6-S347 (5′-TGGATGTCTCACACAATCGG) | mTLR6-AS790 (5′-TCAATAAGGTTGGACCTCTGG) | 444 |
| mTLR-7 | mTLR7-S499 (5′-CATCTTCTGAGCCTTGAGGC) | mTLR7-AS866 (5′-CACGGTGTACACGGATATGG) | 368 |
| mTLR-8 | mTLR8-S941 (5′-AGGACGATTCCTTCTACCTGG) | mTLR8-AS1485 (5′-GCCTTGCCATAAGCAGTACAC) | 545 |
| mTLR-9 | mTLR9-S1195 (5′-CATGAACGGCATCTTCTTCC) | mTLR9-AS1548 (5′-ATAGTCACCAGGTTGTTCCG) | 354 |
a
The primer pairs shown were used in nested amplification reactions for Tlr, MD-2, and CD14 mRNAs. Samples consisted of 5 μl of 1:100 dilutions of the primary RT-PCR products (Table 1; see Materials and Methods). PCR mixes lacking AmpliTaq DNA polymerase were preheated at 94°C for 2 min, and then complete reaction mixes were run as described in Materials and Methods.