Lecture on Delta Delta Ct PCR

Introduction

• Overview of Delta Delta Ct method in PCR
  • Commonly used in gene expression studies
  • Helps in comparing gene expression levels between samples

Basics of PCR

• Polymerase Chain Reaction (PCR) is a technique used to amplify DNA
• Ct (Cycle threshold) is a value in quantitative PCR that represents the cycle number at which the fluorescence exceeds the threshold, indicating the presence of the target gene

Delta Ct Method

• Delta Ct (ΔCt) calculation:
  • ΔCt = Ct (gene of interest) - Ct (reference gene)
  • Normalizes the expression of target genes to a reference gene

Delta Delta Ct (ΔΔCt) Method

• ΔΔCt calculation:
  • ΔΔCt = ΔCt (experiment) - ΔCt (control)
  • Allows for the comparison of gene expression levels between experimental and control groups

Applications

• Useful in comparative gene expression analysis
• Widely employed in studies involving changes in gene expression under different experimental conditions

Advantages

• Simplifies the comparison of gene expression
• Reduces variability by normalizing to a reference gene

Considerations

• Selection of appropriate reference gene is crucial
• Reference gene should have stable expression across samples

Conclusion

• Delta Delta Ct is a powerful method for analyzing relative gene expression
• Essential for studies in molecular biology and genetics