Qingyao Wang
Institute of Molecular Medicine, Huaqiao University, 362021,Quanzhou, China
Huachun Rao
Institute of Molecular Medicine, Huaqiao University, 362021,Quanzhou, China
Huiyong Yang
Institute of Molecular Medicine, Huaqiao University, 362021,Quanzhou, China
Yong Diao
Institute of Molecular Medicine, Huaqiao University, 362021,Quanzhou, China
ABSTRACT
Tri-primer PCR has been developed into a sensitive and specific method for the genotyping of Single Nucleotide Polymorphism (SNP). However, the prior purification of genomic DNA from blood is necessary and time-consuming. In the current study, a special reaction system that leaves out the DNA extraction was investigated. Three different anticoagulant treatment, EDTA, heparin and sodium citrate, all allowed successfully amplication and genotyping directly from blood and filter paper. Spotted on filter paper was less difficult for amplification than crude blood. The methodology described facilitates genotyping with the advantages of immediate sample testing, lower experimental costs and time-saving.
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How to cite this article
Qingyao Wang, Huachun Rao, Huiyong Yang and Yong Diao, 2013. Single Nucleotide Polymorphism Genotyping Directly from Whole Blood and Filter Paper by Improved Allele Specific PCR. Biotechnology, 12: 113-116.
DOI: 10.3923/biotech.2013.113.116
URL: https://scialert.net/abstract/?doi=biotech.2013.113.116
DOI: 10.3923/biotech.2013.113.116
URL: https://scialert.net/abstract/?doi=biotech.2013.113.116
REFERENCES
- Al-Soud, W.A., L.J. Jonsson and P. Radstrom, 2000. Identification and characterization of immunoglobulin G in blood as a major inhibitor of diagnostic PCR. J. Clin. Microbiol., 38: 345-350.
Direct Link - Al-Soud, W.A. and P. Radstrom, 2001. Purification and characterization of PCR-inhibitory components in blood cells. J. Clin. Microbiol., 39: 485-493.
CrossRefDirect Link - Aubry, M., C. Roche, M. Dupont-Rouzeyrol, J. Aaskov and J. Viallon et al., 2012. Use of serum and blood samples on filter paper to improve the surveillance of dengue in Pacific Island Countries. J. Clin. Virol., 55: 23-29.
CrossRefDirect Link - Ar'ev, A.L., N.A. Kunitskaia and L.S. Kozina, 2011. Gout and hyperuricemia today: Prevalence, risk factors, features in the elderly. Adv. Gerontol., 25: 540-544.
Direct Link - Bu, Y., H. Huang and G. Zhou, 2008. Direct Polymerase Chain Reaction (PCR) from human whole blood and Filter-paper-dried blood by using a PCR buffer with a higher pH. Anal. Biochem., 375: 370-372.
CrossRefDirect Link - Choi, H.K., K. Atkinson, E.W. Karlson and G. Curhan, 2005. Obesity, weight change, hypertension, diuretic use and risk of gout in men: The health professionals Follow-up study. Arch. Internal Med., 165: 742-748.
Direct Link - Horakova, H., I. Polakovicova, G.M. Shaik, J. Eitler, V. Bugajev, L. Draberova and P. Draber, 2011. 1, 2-propanediol-trehalose mixture as a potent quantitative real-time PCR enhancer. BMC Biotechnol., Vol. 11.
CrossRef - Koizumi, M., K. Morita, M. Takagi, H. Yasumo and A. Kasuya, 2005. Improvement of single nucleotide polymorphism genotyping by allele-specific PCR using primers modified with an ENA residue. Anal. Biochem., 340: 287-294.
CrossRefDirect Link - Merriman, T.R. and N. Dalbeth, 2011. The genetic basis of hyperuricaemia and gout. Joint Bone Spine, 78: 35-40.
CrossRefDirect Link - Reimer, R.J. and R.H. Edwards, 2004. Organic anion transport is the primary function of the SLC17/type I phosphate transporter family. Pflugers Arch., 447: 629-635.
CrossRefDirect Link - Sharma, R., A.S. Virdi and P. Singh, 2012. A novel method for whole blood PCR without pretreatment. Gene, 501: 85-88.
CrossRefDirect Link - Yang, H.Y., C.C. Xu and G.H. Zhou, 2011. Biological response fingerprinting (BioReF) approach for genome-wide selection of control of herbal medicine formulations. Proceedings of the 7th International Forum on Post-Genome Technologies and China-Japan-Korea Summit on Natural Products, October 27-29, 2011, Chongqing, China, pp: 310.
- You, Y.Q., Q.Y. Wang, C.C. Xu, H.Y. Yang and Y.N. Li, 2013. Association of single nucleotide polymorphisms of SLC2A9, SLC17A3 and ABCG2 gene with gout susceptibility in Quanzhou residents. Chinese J. Rheumatol., 17: 114-118.
Direct Link - Zhang, Z., M.B. Kermekchiev and W.M. Barnes, 2010. Direct DNA amplification from crude clinical samples using a PCR enhancer cocktail and novel mutants of Taq. J. Mol. Diagn., 12: 152-161.
CrossRefDirect Link