A novel multiplex tetra-primer ARMS-PCR for the simultaneous genotyping of six single nucleotide polymorphisms associated with female cancers.
A novel multiplex tetra-primer ARMS-PCR for the simultaneous genotyping of six single nucleotide polymorphisms associated with female cancers.
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DOI：
10.1371/journal.pone.0062126
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Ma X
中科院分区：
文献类型：
--
作者：
Zhang C;Liu Y;Ring BZ;Nie K;Yang M;Wang M;Shen H;Wu X;Ma X
四引物扩增难突变系统PCR （T-ARMS-PCR）是一种快速、经济的分析SNP的方法，只需要PCR扩增和随后的电泳就可以确定基因型。为了提高T-ARMS-PCR的通量和效率，我们将T-ARMS-PCR与基于嵌合引物的温度开关PCR （TSP）策略相结合，并利用毛细管电泳（CE）对扩增子进行分离和鉴定。我们通过同时对四个乳腺癌和两个宫颈癌风险相关snp进行基因分型来评估这一过程。共收集24条T-ARMS-PCR引物，每条引物5 '标记一个通用序列和一对通用引物，扩增186份对照女性血样中6个snp的12个目标等位基因。还对所有样品进行直接测序，以评估该方法的准确性。在186个样本中，单次PCR可产生多达11个扩增子，并通过CE分离。多重T-ARMS-PCR分型结果与所有样品的直接测序结果完全一致。这种新颖的多重T-ARMS-PCR方法是首次报道的方法，可以在一次反应中对6个snp进行基因分型，而不需要除电泳外的pcr后处理。该方法可靠、快速、简便。
The tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) is a fast and economical means of assaying SNP's, requiring only PCR amplification and subsequent electrophoresis for the determination of genotypes. To improve the throughput and efficiency of T-ARMS-PCR, we combined T-ARMS-PCR with a chimeric primer-based temperature switch PCR (TSP) strategy, and used capillary electrophoresis (CE) for amplicon separation and identification. We assessed this process in the simultaneous genotyping of four breast cancer–and two cervical cancer risk–related SNPs. A total of 24 T-ARMS-PCR primers, each 5′-tagged with a universal sequence and a pair of universal primers, were pooled together to amplify the 12 target alleles of 6 SNPs in 186 control female blood samples. Direct sequencing of all samples was also performed to assess the accuracy of this method. Of the 186 samples, as many as 11 amplicons can be produced in one single PCR and separated by CE. Genotyping results of the multiplex T-ARMS-PCR were in complete agreement with direct sequencing of all samples. This novel multiplex T-ARMS-PCR method is the first reported method allowing one to genotype six SNPs in a single reaction with no post-PCR treatment other than electrophoresis. This method is reliable, fast, and easy to perform.
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影响因子：
4.5
作者：
Long J;Cai Q;Shu XO;Qu S;Li C;Zheng Y;Gu K;Wang W;Xiang YB;Cheng J;Chen K;Zhang L;Zheng H;Shen CY;Huang CS;Hou MF;Shen H;Hu Z;Wang F;Deming SL;Kelley MC;Shrubsole MJ;Khoo US;Chan KY;Chan SY;Haiman CA;Henderson BE;Le Marchand L;Iwasaki M;Kasuga Y;Tsugane S;Matsuo K;Tajima K;Iwata H;Huang B;Shi J;Li G;Wen W;Gao YT;Lu W;Zheng W
通讯作者：
Zheng W
影响因子：
2.7
作者：
Dupont, M;Goldsborough, A;Lavabre-Bertrand, T
通讯作者：
Lavabre-Bertrand, T
DOI：
10.1007/978-1-61737-954-3_5
发表时间：
2011-01-01
期刊：
DISEASE GENE IDENTIFICATION: METHODS AND PROTOCOLS
影响因子：
--
作者：
Millis, Meredith P.
通讯作者：
Millis, Meredith P.
影响因子：
8.8
作者：
Harlid, S.;Ivarsson, M. I. L.;Carlson, J.
通讯作者：
Carlson, J.
影响因子：
3.1
作者：
Beck, O;Seidl, C;Koehl, U
通讯作者：
Koehl, U