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Table 1 Primers and PCR conditions.

From: Inhibitory effects of rat bone marrow-derived dendritic cells on naïve and alloantigen-specific CD4+ T cells: a comparison between dendritic cells generated with GM-CSF plus IL-4 and dendritic cells generated with GM-CSF plus IL-10

Primer Sequence (5'- > 3') PCR product Annealing temperature
GAPDH For: GGT CGG TGT GAA CGG ATT TG
Rev: GTG AGC CCC AGC CTT CTC CAT
319 bp 62°C
MHC II For: CAG GAT CTG GAA GGT CCA
Rev: AGC TGT GGT TGT GCT GA
517 bp 55°C
CD40 For: CGC TAT GGG GCT GCT TGT TGA CAG;
Rev: GAC GGT ATC AGT GGT CTC AGT GGC
401 bp 58°C
CD80 For: TGG TGA AAC ACC TGA CCA
Rev: GTT TCT CTG CTT GCC TCA
517 bp 50°C
CD86 For: TGG GAA ACA GAG CTC TCA
Rev: AGG TTG ATC GAC TCG TCA
518 bp 53°C
  1. Primers specific for rat IL-12p35 was purchased from Invitrogen/Biosource. Total RNA was isolated from 106 cells with 1 ml of the ready-to-use reagent Trizol Reagent (Invitrogen GmbH, Germany) according to the manufacturer's recommendations. Reverse transcription of 5 μl RNA was performed using the GeneAmp RNA-PCR-Kit (Applied Biosystems GmbH, Germany). Five μl cDNA was amplified using Gold AmpliTaq DNA-Polymerase (0.05 U/μl) and the specific primers (5 μmol/L each) mixed in nuclease free water (Promega GmbH, Germany) to an end volume of 50 μl per sample.