Multiplex amplification of all coding sequences within 10 cancer genes by Gene-Collector.
Multiplex amplification of all coding sequences within 10 cancer genes by Gene-Collector.
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DOI：
10.1093/nar/gkm078
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发表时间：
2007
影响因子：
14.9
通讯作者：
Davis RW
中科院分区：
文献类型：
--
作者：
Fredriksson S;Banér J;Dahl F;Chu A;Ji H;Welch K;Davis RW
在此，我们提出了基因收集器，用于核酸的多重扩增的方法。该方法已被用于成功地扩增10个人类癌症基因的编码序列，在一个测定与均匀丰度的最终产品。在这种情况下，通过多重PCR用170个引物对启动扩增。然后通过在能够仅并置完全匹配的同源引物对的收集器探针上连接，使每个PCR产物特异性环化。通常与源自使用许多引物对的多重PCR相关的任何扩增伪影，例如假扩增子、引物二聚体等，不会被核酸外切酶处理环化和降解。然后通过随机引发的滚环复制进一步富集环状DNA分子。如通过与定制重测序DNA微阵列杂交所见，90%的靶向扩增子扩增成功。实时荧光定量PCR结果显示，96%的扩增产物均在平均丰度的4倍以内。Gene-Collector可用于许多应用，如高通量重测序、SNP分析和病原体检测。
Herein we present Gene-Collector, a method for multiplex amplification of nucleic acids. The procedure has been employed to successfully amplify the coding sequence of 10 human cancer genes in one assay with uniform abundance of the final products. Amplification is initiated by a multiplex PCR in this case with 170 primer pairs. Each PCR product is then specifically circularized by ligation on a Collector probe capable of juxtapositioning only the perfectly matched cognate primer pairs. Any amplification artifacts typically associated with multiplex PCR derived from the use of many primer pairs such as false amplicons, primer-dimers etc. are not circularized and degraded by exonuclease treatment. Circular DNA molecules are then further enriched by randomly primed rolling circle replication. Amplification was successful for 90% of the targeted amplicons as seen by hybridization to a custom resequencing DNA micro-array. Real-time quantitative PCR revealed that 96% of the amplification products were all within 4-fold of the average abundance. Gene-Collector has utility for numerous applications such as high throughput resequencing, SNP analyses, and pathogen detection.
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影响因子：
7
作者：
Pastinen, T;Raitio, M;Syvänen, AC
通讯作者：
Syvänen, AC
影响因子：
30.8
作者：
Syvänen, AC
通讯作者：
Syvänen, AC
多重PCR：使用热稳定的热嗜热RECA蛋白来最大程度地减少非特异性PCR产物。
影响因子：
14.9
作者：
Shigemori, Yasushi;Mikawa, Tsutomu;Shibata, Takehiko;Oishi, Michio
通讯作者：
Oishi, Michio
影响因子：
14.9
作者：
CHAMBERLAIN, JS;GIBBS, RA;CASKEY, CT
通讯作者：
CASKEY, CT
影响因子：
14.9
作者：
Dahl F;Gullberg M;Stenberg J;Landegren U;Nilsson M
通讯作者：
Nilsson M