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A sticky end-driven PAM-free RPA-CRISPR/Cas12a dual amplification system for ultrasensitive detection of KRAS G12C
Highly sensitive detection of KRAS G12C mutation using a dual amplification system without PAM requirement
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Abstract
The method achieved a detection limit of 1.5 aM for the KRAS G12C mutation.
- A fluorescent biosensing platform was developed to detect a specific genetic mutation without the need for a PAM site.
- The platform utilizes CRISPR/Cas12a combined with a specific DNA amplification technique.
- Detection of KRAS G12C mutations was effective across a linear range from 10 aM to 10 pM.
- The system could identify mutations at a level as low as 0.1% in a sample concentration of 10 pM.
- It demonstrated strong performance in analyzing real biological samples.
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