Selected article for: "High Capacity cdna and real time"
Author: Hernandez, Nicholas; Melki, Isabelle; Jing, Huie; Habib, Tanwir; Huang, Susie S.Y.; Danielson, Jeffrey; Kula, Tomasz; Drutman, Scott; Belkaya, Serkan; Rattina, Vimel; Lorenzo-Diaz, Lazaro; Boulai, Anais; Rose, Yoann; Kitabayashi, Naoki; Rodero, Mathieu P.; Dumaine, Cecile; Blanche, Stéphane; Lebras, Marie-Noëlle; Leung, Man Chun; Mathew, Lisa Sara; Boisson, Bertrand; Zhang, Shen-Ying; Boisson-Dupuis, Stephanie; Giliani, Silvia; Chaussabel, Damien; Notarangelo, Luigi D.; Elledge, Stephen J.; Ciancanelli, Michael J.; Abel, Laurent; Zhang, Qian; Marr, Nico; Crow, Yanick J.; Su, Helen C.; Casanova, Jean-Laurent
Title: Life-threatening influenza pneumonitis in a child with inherited IRF9 deficiency
  • Document date: 2018_10_1
    • ID: jqv0lyfx_45
    Snippet: qRT-PCR Total RNA from PBMCs was extracted using RNAqueous-Micro Kit (Ambio). Reverse transcription was performed using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Messenger RNAs were quantified with IRF9 probes Hs00960976-m1 (exon 1-2) and Hs00196051-m1 (exon 7-8; Thermo Fischer Scientific) by qRT-PCR using the Taqman Gene Expression Assay (Applied Biosystems) and normalized to the expression level of HPRT1. RNA was is.....
    Document: qRT-PCR Total RNA from PBMCs was extracted using RNAqueous-Micro Kit (Ambio). Reverse transcription was performed using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Messenger RNAs were quantified with IRF9 probes Hs00960976-m1 (exon 1-2) and Hs00196051-m1 (exon 7-8; Thermo Fischer Scientific) by qRT-PCR using the Taqman Gene Expression Assay (Applied Biosystems) and normalized to the expression level of HPRT1. RNA was isolated from SV40-fibroblasts, and B-LCL cells were stimulated with 1,000 IU/ml IFN-α2b, IFNβ, or IFN-γ for 2 or 8 h with RNA lysis buffer, treated with DNase, and purified according to the manufacturer's protocol (Zymo Research). RT-PCR was performed using random hexamers and the Superscript III reverse strand synthesis kit according to the manufacturer's instructions (Thermo Fisher Scientific). Quantitative real-time PCR was performed with Applied Biosystems Taqman assays using the β-glucuronidase housekeeping gene for normalization. Results are expressed using the ΔΔCt method, as described by the manufacturer.

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