Table 1.
Primers and parameters for PCR sequencing
| Gene | Primer sequence (from 5′ to 3′) | Parameters of PCR | Restriction enzymes |
|---|---|---|---|
| CRP |
F: GATCTGTGTGATCTGAGAAACCTCCT R: GAGGTACCAGAGACAGAGACGTG |
95°5′; 30 cycles (94°30″, 62°1′, 72°30″), 72°10′ | MaeIII |
| CCR2 |
F: GAAAGTGGATTGAACAAGGA R: CAGGTTGAGCAGGTAAATGT |
95°5′; 30 cycles (94°30″, 58°30″, 72°30″), 72°10′ | FokI |
| CD14 |
F: GTGCCAACAGATGAGGTTCA R: CGCAGCGGAAATCTTCATC |
95°5′; 30 cycles (94°30″, 58°30″, 72°30″), 72°10′ | HaeIII |
| TGF-β |
F: TCGCGGGTGCTGTTGTACA R: TTCAAGACCACCCACCTTCT |
94°5′; 35 cycles (94°30″, 58°30″, 72°30″), 72°10′ | FseI |
| TLR2 |
F: GCCTACTGGGTGGAGAACCT R: GGCCACTCCAGGTAGGTCTT |
95°10′; 35 cycles (94°30″, 60°30″, 72°30″), 72°10′ | AciI |
| TLR4 (1) |
F: GATTAGCATACTTAGACTACTACCTCCATG R: GATCAACTTCTGAAAAAGCATTCCCAC |
95°4′; 30 cycles (95°30″, 55°30″, 72°30″), 72°10′ | NcoI |
| TLR4 (2) |
F: GGTTGCTGTTCTGAAAGTGATTTTGGGAGAA R: ACCTGAAGACTGGAGAGTGAGTTAAATGCT |
95°4′; 30 cycles (95°30″, 55°30″, 72°30″), 72°10′ | HinfI |
| IL-6 |
F: CTGATTGGAAACCTTATTAAG R: TGACTTCAGCTTTACTCTTGT |
95°5′; 30 cycles (94°30″, 48°30″, 72°30″), 72°10′ | Hsp92II |
| IL-1ra |
F: CTCAGCAACACTCCTAT R: TCCTGGTCTGCAAGGTAA |
95°5′; 30 cycles (94°30″, 60°30″, 72°30″), 72°10′ | – |
| 16S RNA |
F: AGTTTGATCCTGGCTCAG R: GGACTACCAGGGTATCTAAT |
95°15′; 35 cycles (60°45″, 72°10″, 95°45″, 60°45″, 72°60″), 72°10′ | – |
F forward, R reverse