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Article

Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously

  • Authors:
    • Rui Yang
    • Jun-He Zhang
    • Guo-Yan Yuan
  • View Affiliations / Copyright

    Affiliations: Department of Life Science and Technology, Xinxiang Medical University, Xinxiang, Henan 453003, P.R. China, Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang, Henan 453003, P.R. China, Department of Neurosurgery, The First Affiliated Hospital of Xinxiang Medical University, Weihui, Henan 453100, P.R. China
  • Pages: 201-204
    |
    Published online on: October 9, 2012
       https://doi.org/10.3892/mmr.2012.1122
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Abstract

The present study aimed to explore the factors that affect polymerase chain reaction using sequence-specific primers (PCR-SSP) and to establish an optimized PCR-SSP method for detecting multiple gene polymorphisms simultaneously. The amplification system parameters, including the concentrations of Mg2+, dNTPs, pfu Taq, primers and control primers, were optimized using the designed PCR-SSP reactions. The resulting optimized reaction system was used to determine the melting temperature of the genomic DNA and the cycling parameters. The optimized PCR-SSP method was used to analyze the polymorphisms of the following genes: mutations -308A/G and -238G/A in TNF-α, -174G/C in IL-6 and C/T mutation at exon 188 of CYP2D6 *10B. The PCR-SSP amplification system was optimized; in a 20 µl reaction system, the quantities of Mg2+, dNTPs, pfu Taq, primers, control primers and genomic DNA were 3.25 µM, 0.5 mM, 2.5 units, 0.5 µM, 0.2 µM and 0.15 µg, respectively. The cycling system comprised 5 start cycles and took 15 min to melt a genomic DNA sample using a touchdown protocol. The optimized PCR-SSP is suitable for polymorphism analysis of polygenic SNPs in large genomic DNA samples and a number of different genes.
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1 

Knight JC: Functional implications of genetic variation in non-coding DNA for disease susceptibility and gene regulation. Clin Sci (London). 104:493–501. 2003. View Article : Google Scholar : PubMed/NCBI

2 

He JQ, Chan-Yeung M, Becker AB, et al: Genetic variants of the IL13 and IL4 genes and atopic diseases in at-risk children. Genes Immun. 4:385–389. 2003. View Article : Google Scholar : PubMed/NCBI

3 

Zhang Y, Zhang J, Tian C, et al: The -308 G/A polymorphism in TNF-α gene is associated with asthma risk: an update by meta-analysis. J Clin Immunol. 31:174–185. 2011.

4 

Faltraco F, Bürger K, Zill P, et al: Interleukin-6–174 G/C promoter gene polymorphism C allele reduces Alzheimer’s disease risk. J Am Geriatr Soc. 51:578–579. 2003.

5 

Licastro F, Grimaldi LM, Bonafé M, et al: Interleukin-6 gene alleles affect the risk of Alzheimer’s disease and levels of the cytokine in blood and brain. Neurobiol Aging. 24:921–926. 2003.

6 

Eley TC, Tahir E, Angleitner A, et al: Association analysis of MAOA and COMT with neuroticism assessed by peers. Am J Med Genet B Neuropsychiatr Genet. 120B:90–96. 2003. View Article : Google Scholar : PubMed/NCBI

7 

Roh HK, Kim CE, Chung WG, Park CS, Svensson JO and Bertilsson L: Risperidone metabolism in relation to CYP2D6*10 allele in Korean schizophrenic patients. Eur J Clin Pharmacol. 57:671–675. 2001.

8 

He JQ, Foreman MG, Shumansky K, et al: Associations of IL6 polymorphisms with lung function decline and COPD. Thorax. 64:698–704. 2009. View Article : Google Scholar : PubMed/NCBI

9 

Córdoba-Lanús E, de-Torres JP, López-Aguilar C, et al: Association of IL-6 gene polymorphisms and COPD in a Spanish population. Resp Med. 102:1805–1811. 2008.

10 

Elmas E, Bugert P, Popp T, et al: The P-selectin gene polymorphism Val168Met: a novel risk marker for the occurrence of primary ventricular fibrillation during acute myocardial infarction. J Cardiovasc Electrophysiol. 21:1260–1265. 2010. View Article : Google Scholar

11 

Fugger L, Morling N, Ryder LP, Odum N and Svejgaard A: Technical aspects of typing for HLA-DP alleles using allele-specific DNA in vitro amplification and sequence-specific oligonucleotide probes. Detection of single base mismatches. J Immunol Methods. 129:175–185. 1990. View Article : Google Scholar

12 

Bon MA, van Oeveren-Dybicz A and van den Bergh FA: Genotyping of HLA-B27 by real-time PCR without hybridization probes. Clin Chem. 46:1000–1002. 2000.PubMed/NCBI

13 

Yang R, Yang X, Qian C and Liu J: Rapid detection of gene polymorphism of clinical pediatrics diseases by PCR-SSP. J Appl Clin Pediatrics. 21:687–688. 2006.(In Chinese).

14 

Schäffler A, Weigert J, Neumeier M, Schölmerich J and Buechler C: Regulation and function of collagenous repeat containing sequence of 26-kDa protein gene product ‘cartonectin’. Obesity (Silver Spring). 15:303–313. 2007.

15 

Grottkau BE, Purudappa PP and Lin YF: Multilineage differentiation of dental pulp stem cells from green fluorescent protein transgenic mice. Int J Oral Sci. 2:21–27. 2010. View Article : Google Scholar : PubMed/NCBI

16 

Aldener-Cannavá A and Olerup O: HLA-DOB1 “low-resolution” typing by PCR amplification with sequence-specific primers (PCR-SSP). Eur J Immunogenet. 21:447–455. 1994.

17 

Zetterquist H and Olerup O: Identification of the HLA-DRB1*04, -DRB1*07, and -DRB1*09 alleles by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours. Hum Immunol. 34:64–74. 1992.

18 

Olerup O and Zetterquist H: HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: an alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation. Tissue Antigens. 39:225–235. 1992. View Article : Google Scholar

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Copy and paste a formatted citation
Spandidos Publications style
Yang R, Zhang J and Yuan G: Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously. Mol Med Rep 7: 201-204, 2013.
APA
Yang, R., Zhang, J., & Yuan, G. (2013). Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously. Molecular Medicine Reports, 7, 201-204. https://doi.org/10.3892/mmr.2012.1122
MLA
Yang, R., Zhang, J., Yuan, G."Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously". Molecular Medicine Reports 7.1 (2013): 201-204.
Chicago
Yang, R., Zhang, J., Yuan, G."Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously". Molecular Medicine Reports 7, no. 1 (2013): 201-204. https://doi.org/10.3892/mmr.2012.1122
Copy and paste a formatted citation
x
Spandidos Publications style
Yang R, Zhang J and Yuan G: Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously. Mol Med Rep 7: 201-204, 2013.
APA
Yang, R., Zhang, J., & Yuan, G. (2013). Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously. Molecular Medicine Reports, 7, 201-204. https://doi.org/10.3892/mmr.2012.1122
MLA
Yang, R., Zhang, J., Yuan, G."Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously". Molecular Medicine Reports 7.1 (2013): 201-204.
Chicago
Yang, R., Zhang, J., Yuan, G."Establishment of an optimized PCR method using sequence-specific primers for screening multiple gene polymorphisms simultaneously". Molecular Medicine Reports 7, no. 1 (2013): 201-204. https://doi.org/10.3892/mmr.2012.1122
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