Geneamp 5700 sequence detection system software

Total RNA was extracted from decidua with the Rneasy Mini Kits (QIAGEN, Valencia, CA) according to the manufacturer’s instructions. For qRT-PCR, amplification was performed in an ABI5700 (PE Biosystems, Foster City, CA) using the SYBR Green kit (QuantiTect SYBR Green PCR; QIAGEN). The primers were designed from the target human mRNA sequence using Primer Express software (Applied Biosystems). Each primer was entered into an NCBI BLAST search to ensure that it was specific for the target mRNA transcription. The primer sequences are the following: IL-17 forward 5’-CCG GAC TGT GAT GGT CAA-3′, reverse 5′- CTC ATT GCG GTG GAG ATT-3′; IL-10 forward 5’-GAC TTT AAG GGT TAC CTG GGT TG-3′, reverse 5’-TCA CAT GCG CCT TGA TGT CTG-3′; TGF-β1 forward 5’-CAA TTC CTG GCG ATA CCT CAG, reverse 5’-GCA CAA CTC CGG TGA CAT CAA-3′. The housekeeping gene β-actin primers, forward 5′- ACG TTG CTA TCC AGG CTG TGC TAT-3′, and reverse 5′-TTA ATG TCA CGC ACG ATT TCC CGC-3′ were used with all samples. The primers were synthesized by BioAsia Co. (Shanghai, China). The cycling conditions were 15 s at 95 °C, 45 cycles of 5 s at 95 °C, 20s at 60 °C, 10s at 72 °C, and 15 s at 65 °C. Data were analyzed using the GeneAmp 5700 Sequence Detection System software (version 1.1; Applied Biosystems, Foster City, CA) and were converted into threshold cycle (Ct) values.