Selected article for: "gene expression and quantitative RT PCR"
Author: Chang, Stewart T.; Sova, Pavel; Peng, Xinxia; Weiss, Jeffrey; Law, G. Lynn; Palermo, Robert E.; Katze, Michael G.
Title: Next-Generation Sequencing Reveals HIV-1-Mediated Suppression of T Cell Activation and RNA Processing and Regulation of Noncoding RNA Expression in a CD4(+) T Cell Line
  • Document date: 2011_9_20
    • ID: zyzgk2z3_36
    Snippet: Quantitative reverse transcription (RT-PCR). RNA was reverse transcribed using the QuantiTect reverse transcription kit (Qiagen, Valencia, CA). The resulting cDNA samples were diluted 50ϫ. ABI TaqMan assays were run for each sample in triplicate (see Fig. S3C in the supplemental material). Relative expression was calculated using the ⌬⌬C T method with averaged ⌬C T values (where C T stands for threshold cycle) for the OAZ1 gene as a calibr.....
    Document: Quantitative reverse transcription (RT-PCR). RNA was reverse transcribed using the QuantiTect reverse transcription kit (Qiagen, Valencia, CA). The resulting cDNA samples were diluted 50ϫ. ABI TaqMan assays were run for each sample in triplicate (see Fig. S3C in the supplemental material). Relative expression was calculated using the ⌬⌬C T method with averaged ⌬C T values (where C T stands for threshold cycle) for the OAZ1 gene as a calibrator, as the expression of OAZ1 did not significantly change between 12 and 24 hpi in the NGS data. Intracellular viral RNA load was quantified as previously described (45) . Relative change of IRF1 was determined by qPCR using primers PSK1 (forward primer [5= TCTG GCTTTTTCCTCTGAGC 3=] and reverse primer [5= ATGCTTTTCTGG GGTCACTG 3=]).

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