Selected article for: "endogenous control and real time"
Author: van Zuylen, Wendy J.; Doyon, Priscilla; Clément, Jean-François; Khan, Kashif Aziz; D'Ambrosio, Lisa M.; Dô, Florence; St-Amant-Verret, Myriam; Wissanji, Tasheen; Emery, Gregory; Gingras, Anne-Claude; Meloche, Sylvain; Servant, Marc J.
Title: Proteomic Profiling of the TRAF3 Interactome Network Reveals a New Role for the ER-to-Golgi Transport Compartments in Innate Immunity
  • Document date: 2012_7_5
    • ID: 1m5dbwjv_42
    Snippet: After stimulation, total RNA was extracted from HeLa cells using Trizol reagent (Invitrogen). 2 mg of RNA was reverse transcribed using the High Capacity cDNA Reverse Transcription Kit with random primers (Applied Biosystems) as described by the manufacturer. SYBR green PCR reactions were performed using 2 ml of cDNA samples (25-50 ng), 5 ml of the Fast SYBR qPCR Master Mix (Applied Biosystems) and 10 pmol of each primer in a total volume of 10 m.....
    Document: After stimulation, total RNA was extracted from HeLa cells using Trizol reagent (Invitrogen). 2 mg of RNA was reverse transcribed using the High Capacity cDNA Reverse Transcription Kit with random primers (Applied Biosystems) as described by the manufacturer. SYBR green PCR reactions were performed using 2 ml of cDNA samples (25-50 ng), 5 ml of the Fast SYBR qPCR Master Mix (Applied Biosystems) and 10 pmol of each primer in a total volume of 10 ml. The IFN qRT-Primer set for real-time quantification of the IFN response (IFNb, ISG56 (ifit1) and OAS1) was purchased from InvivoGen (San Diego, CA). The ABI PRISM 7900HT Sequence Detection System (Applied Biosystems) was used to measure the amplification level. All reactions were run in triplicate and the average Cts were used for quantification. TBP (TATA binding protein) was used as endogenous control.

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