ICCAS OpenIR
Single-nucleotide polymorphism (SNP) genotyping using cationic conjugated polymers in homogeneous solution
Duan, Xinrui1; Yue, Wei2; Liu, Libing1; Li, Zhengping3; Li, Yuliang1; He, Fuchu2; Zhu, Daoben1; Zhou, Gangqiao2; Wang, Shu1
2009
Source PublicationNATURE PROTOCOLS
ISSN1754-2189
Volume4Issue:6Pages:984-991
AbstractThis protocol describes a simple, convenient and sensitive single-nucleotide polymorphism (SNP) genotyping method using an optically amplifying cationic conjugated polymer and single base primer extension reaction. The fluorescence resonance energy transfer (FRET) efficiency between the conjugated polymer (PFP, poly{(1,4-phenylene)-2,7-[9,9-bis(6'-N, N, N-trimethyl ammonium)-hexyl fluorene] dibromide}) and a fluorescein-labeled dNTP (dNTP-Fl) is correlated to the incorporation of the dNTP-Fl into an allele-specific primer; incorporation occurs by a single base extension reaction when the target DNA and the primer are complementary at the SNP site. By triggering the FRET from PFP to fluorescein and measuring the change in fluorescence intensity of samples, the SNP genotypes can be discriminated. In comparison with other SNP genotyping methods, this protocol simplifies procedures and improves sensitivity by eliminating the need for primer labeling, cumbersome workups, chemical/enzymatic coupling reactions and sophisticated instruments. The assay takes about 2 h for PCR amplification followed by 5.5-7.5 h to obtain the genotypes.
DOI10.1038/nprot.2009.70
Indexed BySCI
Language英语
WOS IDWOS:000284097100002
PublisherNATURE PUBLISHING GROUP
Citation statistics
Cited Times:34[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.iccas.ac.cn/handle/121111/65890
Collection中国科学院化学研究所
Corresponding AuthorWang, Shu
Affiliation1.Chinese Acad Sci, Inst Chem, Beijing Natl Lab Mol Sci, Key Lab Organ Solids, Beijing 100080, Peoples R China
2.Beijing Inst Radiat Med, State Key Lab Prote, Beijing Proteome Res Ctr, Beijing, Peoples R China
3.Hebei Univ, Coll Chem & Environm Sci, Baoding, Hebei Province, Peoples R China
Recommended Citation
GB/T 7714
Duan, Xinrui,Yue, Wei,Liu, Libing,et al. Single-nucleotide polymorphism (SNP) genotyping using cationic conjugated polymers in homogeneous solution[J]. NATURE PROTOCOLS,2009,4(6):984-991.
APA Duan, Xinrui.,Yue, Wei.,Liu, Libing.,Li, Zhengping.,Li, Yuliang.,...&Wang, Shu.(2009).Single-nucleotide polymorphism (SNP) genotyping using cationic conjugated polymers in homogeneous solution.NATURE PROTOCOLS,4(6),984-991.
MLA Duan, Xinrui,et al."Single-nucleotide polymorphism (SNP) genotyping using cationic conjugated polymers in homogeneous solution".NATURE PROTOCOLS 4.6(2009):984-991.
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