Selected article for: "cellular gene and gene expression"
Author: Griffiths, Samantha J.; Koegl, Manfred; Boutell, Chris; Zenner, Helen L.; Crump, Colin M.; Pica, Francesca; Gonzalez, Orland; Friedel, Caroline C.; Barry, Gerald; Martin, Kim; Craigon, Marie H.; Chen, Rui; Kaza, Lakshmi N.; Fossum, Even; Fazakerley, John K.; Efstathiou, Stacey; Volpi, Antonio; Zimmer, Ralf; Ghazal, Peter; Haas, Jürgen
Title: A Systematic Analysis of Host Factors Reveals a Med23-Interferon-? Regulatory Axis against Herpes Simplex Virus Type 1 Replication
  • Document date: 2013_8_8
    • ID: 0lyt8gfq_49
    Snippet: Hela cells were transfected with selected SMARTpool siRNAs in 96-well plates, in triplicate, as described. After 48 h transfection, medium was removed, cells rinsed in PBS and lysed in 100 ml TRIZOL (Invitrogen). Triplicate wells were combined, and RNA extracted by standard phenol:chloroform extraction methods. mRNA levels were determined by TaqMan qPCR, using the one-step RT-qPCR kit (Thermofisher), with gene-specific primers ( Table S9 in Text .....
    Document: Hela cells were transfected with selected SMARTpool siRNAs in 96-well plates, in triplicate, as described. After 48 h transfection, medium was removed, cells rinsed in PBS and lysed in 100 ml TRIZOL (Invitrogen). Triplicate wells were combined, and RNA extracted by standard phenol:chloroform extraction methods. mRNA levels were determined by TaqMan qPCR, using the one-step RT-qPCR kit (Thermofisher), with gene-specific primers ( Table S9 in Text S2), and probes from the Universal Probe Library (Roche). Expression levels normalized to the housekeeping cellular gene hypoxanthine phosphoribosyltransferase 1 (HPRT) and calibrated to mock-transfected cells. qPCR was carried out in duplicate for each sample, and the mean of normalized expression levels calculated.

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