Dewi, Anggi Laksmita and Paramita, Dewi Kartikawati and Facliiroli, Jajah (2021) Development of Tetra-primer Amplification Refractory Mutation System (ARMS) PCR for Detection of CHRNA3 rs8040868. Indonesian Biomedical Journal, 13 (2). 192 – 200. ISSN 20853297
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Abstract
BACKGROUND: Single nucleotide variations (SNV) have been mapped to be associated with several human conditions and diseases. To validate the association between SNV to certain human traits or diseases, a large number of subjects must be included. Thus, in need of a fast, relatively economic, and reliable genotyping method. This can be achieved through the use of tetra-primer amplification refractory mutation system polymerase chain reaction (Tetra-primer ARMS PCR). This study reports strategy to develop Tetra-primer ARMS PCR-based genotyping of CHRNA3 rs8040868. METHODS: The optimization of Tetra-primer ARMS PCR was done through these steps: identification of gene sequence and position of single mutation; designing outer and inner PCR primers; amplification of target gene fragments through PCR by using outer primer; confirming genotype of the PCR product by using sequencing; determining an optimum ratio of outer and inner primer; and determining optimum annealing temperature and cycles for the PCR program. The PCR products were run in 2 gel agarose electrophoresis and visualized under UV illumination. RESULTS: Outer and inner primer ratio of 1:3 with annealing temperature of 64.4oC and 40x cycles was found to be the most optimum condition. Tetra-primer ARMS PCR was able to confirm the results of the DNA sequence of 2 samples, confirming wild-type variants (TT allele) and the heterozygous mutant (CT allele). CONCLUSION: Tetra-primer ARMS PCR was able to genotype rs8040868 of the CHRNA3 gene. © 2021. All Rights Reserved.
Item Type: | Article |
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Additional Information: | Cited by: 1; All Open Access, Gold Open Access |
Uncontrolled Keywords: | tetra-primer ARMS PCR, CHRNA3, rs8040868, genotyping |
Subjects: | R Medicine > RB Biomedical Sciences |
Divisions: | Faculty of Pharmacy |
Depositing User: | Sri JUNANDI |
Date Deposited: | 18 Sep 2024 07:05 |
Last Modified: | 18 Sep 2024 07:05 |
URI: | https://ir.lib.ugm.ac.id/id/eprint/4841 |