Citation: Mou Xian-Bo, Ali Zeeshan, Li Bo, Li Tao-Tao, Yi Huan, Dong Hong-Ming, He Nong-Yue, Deng Yan, Zeng Xin. Multiple genotyping based on multiplex PCR and microarray[J]. Chinese Chemical Letters, ;2016, 27(11): 1661-1665. doi: 10.1016/j.cclet.2016.04.005 shu

Multiple genotyping based on multiplex PCR and microarray

Mou Xian-Bo ^a ,
Ali Zeeshan ^a ,
Li Bo ^a ,
Li Tao-Tao ^a ,
Yi Huan ^a ,
Dong Hong-Ming ^b ,
He Nong-Yue ^a,b,, ,
Deng Yan ^b,, ,
Zeng Xin ^c,,

a.
State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China
b.
Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province, Hunan Key Laboratory of Green Packaging and Application of Biological Nanotechnology, Hunan University of Technology, Zhuzhou 412007, China
c.
Nanjing Maternity and Child Health Care Hospital, Nanjing 210029, China

Corresponding author: He Nong-Yue, nyhe1958@163.com Deng Yan, hndengyan@126.com Zeng Xin, august555482@126.com
Received Date: 15 February 2016
Revised Date: 4 April 2016
Accepted Date: 6 April 2016
Available Online: 20 November 2016

Figures(6)

The genetic variability has obtained more and more attention in the process of diagnosis and treatment of tumors.Herein, we have described a multiple genotyping method based on magnetic enrichmentmultiplex PCR (MEM-PCR) and microarray technology.Monodisperse magnetic beads were fabricated and modified with streptavidin.Four loci on two genes (M235T and A-6G loci on AGT gene, A1298C and C677T loci on MTHFR gene) were selected to study single nucleotide polymorphisms (SNP).Target sequences of these SNP loci were amplified using Cy3-labeled primers through multiplex PCR in one tube after the templates were enriched and purified by functional magnetic beads (MB).Four pairs of NH₂-labeled probes, corresponding to each locus, were fixed on CHO-modified glass slide by covalent binding.Hybridization between target sequences and probes was performed under suitable conditions.The spotting locations on microarray and the ratio of fluorescence intensity, produced by different loci, were used to distinguish the SNP genotypes.Finally, three of gastric cancer samples were collected and genotyping analysis for these four SNP loci was carried out successfully simultaneously by this method.
- Magnetic beads,
- Multiplex PCR,
- Microarray,
- Multiple genotyping,
- Gastric cancer
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