Magnetic Particle-based Single Nucleotide Polymorphism Detection Method Using Linker-Polymerase Chain Reaction

被引：0

作者：

Liu Hongna ^{[1
]}

Li Song ^{[1
,2
]}

Liu Lishang ^{[2
]}

Tian Lan ^{[1
]}

Deng Yan ^{[1
,2
]}

Li Zhiyang ^{[1
]}

Dai Yabin ^{[1
]}

He Nongyue ^{[1
,2
]}

机构：

[1] Southeast Univ, State Key Lab Bioelect, Nanjing 210096, Peoples R China

[2] Hunan Univ Technol, Hunan Key Lab Green Packaging & Applicat Biol Nan, Zhuzhou 412008, Hunan, Peoples R China

来源：

ACTA CHIMICA SINICA | 2009年 / 67卷 / 15期

关键词：

linker-PCR; magnetic particle; dual-color hybridization; SNP type; MULTIPLEX; PCR;

D O I：

暂无

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

An approach was presented to type multiplex single nucleotide polymorphism (SNP) by means of magnetic particles and amplification of whole genome products using universal linker-PCR. The linker-PCR products were captured by magnetic particles modified with streptavidin to fabricate SNP library. Each SNP locus in the library was interrogated by hybridization with a pair of dual-color fluorescence (Cy3, Cy5) probe, and the genotypes could be simultaneously identified by scanning the microarray printed with the denatured fluorescent probes on an unmodified glass slide. This method was applied to analysis the methylenetetrahydrofolate reductase (MTHFR) gene C677T and A 1298C polymorphisms for ten different samples. The fluorescent ratios (match/mismatch signal) of homozygous samples were over 3. This simple and reliable genotyping method is suitable for high-throughput genotyping for multiplex loci, and will find a wide potential application to research and medical diagnosis.

引用

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页码：1797 / 1802

页数：6

共 17 条

[1] Cheng Q, 2004, ACTA CHIM SINICA, V62, P2447
[2] Pharmacogenomics: Translating functional genomics into rational therapeutics
Evans, WE
Relling, MV
[J]. SCIENCE, 1999, 286 (5439) : 487 - 491
[3] Fang CY, 2008, ACTA CHIM SINICA, V66, P2174
[4] Multiplexed genotyping with sequence-tagged molecular inversion probes
Hardenbol, P
Banér, J
Jain, M
Nilsson, M
Namsaraev, EA
Karlin-Neumann, GA
Fakhrai-Rad, H
Ronaghi, M
Willis, TD
Landegren, U
Davis, RW
[J]. NATURE BIOTECHNOLOGY, 2003, 21 (06) : 673 - 678
[5] Hui S, 2007, CHEM J CHINESE U, V28, P1690
[6] Integrated nanoparticle-biomolecule hybrid systems: Synthesis, properties, and applications
Katz, E
Willner, I
[J]. ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2004, 43 (45) : 6042 - 6108
[7] Primer-engineered multiplex PCR-RFLP for detection of MTHFR C677T, prothrombin G20210A and factor V Leiden mutations
Koksal, Vedat
Baris, Ibrahim
Etlik, Ozdal
[J]. EXPERIMENTAL AND MOLECULAR PATHOLOGY, 2007, 83 (01) : 1 - 3
[8] KRUGLYAK L, 1997, NAT GENET, V17, P21224
[9] Li JP, 2008, ACTA CHIM SINICA, V66, P84
[10] 'Playing chicken' over gene markers
Marshall, E
[J]. SCIENCE, 1997, 278 (5346) : 2046 - 2048

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