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Date Published: 28/06/2023
Abstract Views: 341
Views PDF: 258
DOI: https://doi.org/10.56535/jmpm.v48i5.358
Issue
Vol. 48 No. 5 (2023): JOURNAL OF MILITARY PHARMACO-MEDICINE 5-2023
Section
Articles
How to Cite
Nguyễn, L. T., & Bùi, K. C. (2023). APPLICATION OF DROPLET DIGITAL PCR FOR RNA ABSOLUTE QUANTIFICATION. Journal of Military Pharmaco-medicine, 48(5), 30-36. https://doi.org/10.56535/jmpm.v48i5.358

APPLICATION OF DROPLET DIGITAL PCR FOR RNA ABSOLUTE QUANTIFICATION

Nguyen Linh Toan1, Bui Khac Cuong2,
1 Department of Pathophysiology, Vietnam Military Medical University
2 Trung tâm nghiên cứu động vật thực nghiệm, Học viện quân y

Main Article Content

Abstract

Objectives: Droplet Digital PCR (ddPCR) is a PCR method using a microdroplet system that allows the measurement of thousands of independent amplification events in a single reaction. The ddPCR technique requires lower sample and reagent volumes, and reduces overall costs compared to other methods while maintaining high sensitivity and accuracy. Methods: The study was conducted by the experimental method, using ddPCR to quantify the number of RNA copies in research samples. The quantification process on the ddPCR system includes microdroplet generation, amplification, microdroplet reading and data analysis to calculate RNA concentration. Results: RNA concentration in undiluted, diluted samples by a factor of 10 and 100 were 380 copies/µL, 40.7 copies/µL and 3.4 copies/µL, respectively. The signal distribution histogram showed a significant difference in signal intensity between the two populations of microdroplets with and without amplification products. The data showed a very tight positive linear correlation between RNA concentration and the dilution factors. Conclusion: ddPCR is a highly sensitive technique for RNA absolute quantification.

Article Details

Keywords

Droplet Digital PCR, ddPCR, Quantification

References

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