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:: Scientific Journal of Kurdistan University of Medical Sciences- NO 4, 2022 ::
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A review of optimization strategies and the advantages and disadvantages of Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) in genomics studies
Kheirollah Yari 1, Zeinab Jamshidi2
1- , Khirollah.yari@yahoo.com
2- BSc Student, Biotechnology Department, Faculty of Sciences, Malayer University, Malayer, Iran
Abstract:   (66 Views)
Background and Aim: There are several methods for genotyping single nucleotide polymorphisms (SNPs). Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) is an inexpensive, quick, and reliable method that is applicable for SNPs genotyping that saves time and costs compared to other related methods. In the PCR-CTPP technique, allele-specific products of the gene of interest are selectively amplified by adding two pairs of four primers and ordinarily prepared PCR mixture in a single PCR tube. Four allele-specific primers are as follows; F1 (sense) and R1 (antisense) primers for one allele that F1 has an anti-sense base of SNP site at the 3′ end and F2 (sense) and R2 (antisense) for the other allele that F2 has a sense base of SNP site at the 3′ end. PCR-CTPP produces three products with different sizes that make possible genotyping of SNP by gel electrophoresis. To increase the reproducibility and accuracy of the PCR-CTPP results, it is recommended to optimize protocol before sample genotyping. The aim of this study was to review the currently used strategies to optimum application and introduce advantages and disadvantages of the PCR-CTPP method in SNPs genotyping.
 Materials and Methods: The related articles related to the PCR-CTPP method were searched from databases such as ISI Web of science, Science direct, Pubmed, Google Scholar, SID, and Scopus.
Results: For more efficiency and reproducibility of the PCR-CTPP method, it is recommended to pay attention to the specificity and similarity of melting temperatures of the designed primers with specific primer designing algorithms and software.
Conclusion: Strategies for optimizing the concentration ratio of internal and external primers, adding amplification additives to the PCR reaction mix, and using the touchdown program are also suggested.
Keywords: PCR, Polymerase chain reaction with confronting two-pair primers, Genomics, Single Nucleotide Polymorphism, Genotyping
     
Type of Study: Review | Subject: Molecular Medicine and Genetics
Received: 2021/08/24 | Accepted: 2022/01/3
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Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
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مجله علمی دانشگاه علوم پزشکی کردستان Scientific Journal of Kurdistan University of Medical Sciences
مجله علمی دانشگاه علوم پزشکی کردستان Scientific Journal of Kurdistan University of Medical Sciences
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