Rnase h reverse transcription

Supernatants of in vitro cell cultures were analyzed via ELISA using a commercial assay system (eBioScience). For qRT-PCR, total RNA was isolated using TRIzol reagent (Molecular Research Center, Inc.) and subjected to cDNA synthesis using RNase H-reverse transcription (Invitrogen) and oligo (dT) primers. qRT-PCR was performed in triplicate using an iCycler Sequence Detection System (Bio-Rad) and iQTM SYBR Green Supermix (Bio-Rad). The expression of individual genes was calculated with a standard curve and normalized to the expression of Actb. The gene-specific PCR primers (all for mouse genes) are shown in Table 1.

The supernatants of in vitro cell cultures were analyzed by ELISA using a commercial assay system (eBioscience). Autoantibodies for double-stranded DNA and nuclear antigen in the sera collected from 10-mo-old mice were measured using specific ELISA kits (Alpha Diagnostic).
For qRT-PCR, total RNA was isolated using TRIzol reagent (Molecular Research Center) and subjected to cDNA synthesis using RNase H-reverse transcription (Invitrogen) and oligo (dT) primers. qRT-PCR was performed in triplicate using the iCycler Sequence Detection System (Bio-Rad) and iQTM SYBR Green Supermix (Bio-Rad). The expression of individual genes was calculated using a standard curve method and normalized to the expression of Actb. The gene-specific PCR primers (all for mouse genes) are shown in Table S7.