Quantification of Circular RNAs Using Digital Droplet PCR

Area of Science:

• Molecular Biology
• Biochemistry
• Genomics

Background:

• Circular RNAs (circRNAs) are novel regulatory molecules found in body fluids.
• CircRNAs play roles in gene expression and serve as potential disease biomarkers due to their stability.
• Digital droplet PCR (dd-PCR) is a sensitive method for nucleic acid quantification.

Purpose of the Study:

• To detail a workflow for the precise quantification of specific circular RNA (circRNA) levels in cellular samples.
• To demonstrate the application of dd-PCR for accurate circRNA measurement.

Main Methods:

• Design of divergent primers specific for circRNAs.
• RNA extraction and complementary DNA (cDNA) synthesis.
• Digital droplet PCR (dd-PCR) for absolute quantification of circRNAs.

Main Results:

• Achieved precise quantification of target circRNA molecules.
• Demonstrated the utility of dd-PCR for circRNA analysis.

Conclusions:

• dd-PCR enables accurate and sensitive measurement of circRNA levels.
• This methodology supports circRNA research in gene expression and diagnostics.
• The described workflow facilitates robust circRNA quantification in cellular contexts.