Absolute Quantitative Analysis Service of Gene Expression by qPCR

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Absolute Quantitative Analysis

Absolute quantification uses external genes to determine the absolute amount of target DNA(RNA) acid of interest. The ideal external gene contains sequences that are the same as the target sequence. Equivalent amplification efficiencies between the target and external gene are necessary for absolute quantification. Once a suitable construct is identified, a standard curve of external gene dilutions is generated and used to determine the concentrations of unknown target samples.

The main applications of absolute quantitative analysis by qPCR include:
• absolute quantitative analysis of DNA(RNA)
• viral load quantification
• pathogen quantification
• transgene copy determination
• Determination of deactivation ratio of RNAi genes

Absolute quantitative analysis of target gene

Calculation Method of Relative Quantitative Analysis

The amount of the target gene is calculated by using the CT value of real-time PCR and the corresponding standard curve.

Example

Detect the copy number changes of nodavirus after it infect the HighFive cells by absolute quantification qPCR.

Sample description:

Detect the copy numbers after the nodavirus infect HighFive cells for 4 days and 10 days.

Sample names: noda-HF(control), noda-HF(4d),noda-HF(10d).

Detected genes: noda-RNA2

Detection method: Absolute quantification (TaqMan probe qPCR)

Test experiment report:
1. Standard curves of Nodavirus RNA2

Amplification curves of RNA2

Standard curve of RNA2

2. Report charts of original data for detected samples

3. Analysis report of test results

Analysis result of the copy numbers of nodavirus in HighFive cells

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Real-time PCR Service: Gene Expression Analysis by qPCR+
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- Taqman probe qPCR Method
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