Question

Relative gene expression analysis was performed for 3 genes (Caspase-3, Casp-7 and Casp-9) by PCR. Ct values were obtained (data set below) for five patients with a neurodegenerative disease. No other criteria such as age or gender were considered.
Casp-3: 34.56 – 35.67 – 32.19 – 46.43 – 67.23
Casp-7: 57.34 – 46.78 – 75.67- 34.56 – 54.34
Casp-9: 85.67 – 86.45 – 74.56 – 67.59 – 68.67

Answer 1

To analyze the relative gene expression levels of Caspase-3, Caspase-7, and Caspase-9 for the five patients, we need to first calculate the delta Ct values for each gene compared to the housekeeping gene (reference gene).
Assuming that a housekeeping gene was used as a reference and its Ct value is 20 for all samples, we can calculate the delta Ct values for each gene as follows:
Delta Ct (Casp-3) = Ct (Casp-3) - Ct (housekeeping gene)
Delta Ct (Casp-7) = Ct (Casp-7) - Ct (housekeeping gene)
Delta Ct (Casp-9) = Ct (Casp-9) - Ct (housekeeping gene)
For example, for the first patient:
Delta Ct (Casp-3) = 34.56 - 20 = 14.56
Delta Ct (Casp-7) = 57.34 - 20 = 37.34
Delta Ct (Casp-9) = 85.67 - 20 = 65.67
Now, we can compare the relative expression levels of the three genes for each patient. The lower the delta Ct value, the higher the gene expression level.
Patient 1:
Casp-3: 14.56
Casp-7: 37.34
Casp-9: 65.67
Patient 2:
Casp-3: 15.67
Casp-7: 26.78
Casp-9: 66.45
Patient 3:
Casp-3: 12.19
Casp-7: 55.67
Casp-9: 54.56
Patient 4:
Casp-3: 26.43
Casp-7: 14.56
Casp-9: 47.59
Patient 5:
Casp-3: 47.23
Casp-7: 34.34
Casp-9: 48.67
Based on this analysis, we can determine the relative gene expression levels of Caspase-3, Caspase-7, and Caspase-9 for the five patients with the neurodegenerative disease.