Selected article for: "CT threshold cycle and relative expression"
Author: Zapata, Juan Carlos; Carrion, Ricardo; Patterson, Jean L.; Crasta, Oswald; Zhang, Yan; Mani, Sachin; Jett, Marti; Poonia, Bhawna; Djavani, Mahmoud; White, David M.; Lukashevich, Igor S.; Salvato, Maria S.
Title: Transcriptome Analysis of Human Peripheral Blood Mononuclear Cells Exposed to Lassa Virus and to the Attenuated Mopeia/Lassa Reassortant 29 (ML29), a Vaccine Candidate
  • Document date: 2013_9_12
    • ID: 0epeljaf_25
    Snippet: Real-time PCR was performed on an ABI Fast 7900 Real-Time PCR System (Applied Biosystems, Foster City, CA) using the following cycling parameters: 10 min at 95uC (heat activation step); 40 cycles of 15 sec at 95uC, 1 min at 60uC. Relative changes in gene expression were calculated using the DDC t (threshold cycle) method. This method first subtracts the ct (threshold cycle number) of the gene-average ct of the 5 house keeping genes on the array t.....
    Document: Real-time PCR was performed on an ABI Fast 7900 Real-Time PCR System (Applied Biosystems, Foster City, CA) using the following cycling parameters: 10 min at 95uC (heat activation step); 40 cycles of 15 sec at 95uC, 1 min at 60uC. Relative changes in gene expression were calculated using the DDC t (threshold cycle) method. This method first subtracts the ct (threshold cycle number) of the gene-average ct of the 5 house keeping genes on the array to normalize to the RNA amounts. Finally the DDct is calculated which involves subtracting the normalized average ct of LASV samples -normalized average ct of the Control Samples. Then this DDct is raised to the negative power of 2 in order to calculate the fold changes. The p value was calculated using a 2tailed Student's t test. These results were treated to SAM data analysis to avoid donor variation.

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