Total RNA from MEFs was isolated with the RNeasy kit (Qiagen) according to the manufacturer’s instructions and was reverse-transcribed with iScript Reserve Transcription Supermix (Bio-Rad). Ifnb and Il6 mRNA was detected with iQ SYBR Green Supermix (Bio-Rad) and prevalidated primers as described previously (72 (link)). Data were analyzed with the 2ΔΔCt method as described previously (73 (link)).
Iscript reserve transcription supermix
Manufactured by Bio-Rad
The IScript Reserve Transcription Supermix is a reagent designed for reverse transcription of RNA into cDNA. It contains the necessary components for efficient and reliable cDNA synthesis, including reverse transcriptase enzyme, buffer, and stabilizers.
Automatically generated - may contain errors
2 protocols using iscript reserve transcription supermix
1
Quantification of Antiviral Response Genes
2
RNA Isolation and Real-Time PCR Analysis
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Total RNA was isolated using Trizol (Invitrogen). 500 ng of RNA was reverse transcribed to cDNA by using iScript Reserve Transcription Supermix (Bio-Rad). Real time PCR were performed with SYBR green on C1000 CFX96 real time system (Bio-Rad). The primers used are listed in Table S1 .
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