PrimerStation: a highly specific multiplex genomic PCR primer design server for the human genome.

PrimerStation: a highly specific multiplex genomic PCR primer design server for the human genome.
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Priperstation:人类基因组的高度特定的多重基因组PCR底漆设计服务器。

DOI:
10.1093/nar/gkl297
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发表时间:
2006-07-01
影响因子:
14.9
通讯作者:
Morishita S
Morishita S
中科院分区:
生物学2区
文献类型:
--
作者:
Yamada T;Soma H;Morishita S

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PrimerStation () 是一项计算引物集的网络服务,可保证针对整个人类基因组的高特异性。为了达到高精度,我们使用液体溶液中引物的杂交比例。根据严格杂交比率计算序列杂交的状态在计算上成本很高,并且没有网络服务检查整个人类基因组并返回使用精确的物理化学模型计算的高度特异性引物组。为了缩短响应时间,我们提前大约 3 个月使用具有 100 个 CPU(SunFire 15 K)的大规模并行计算机预先计算了特定引物的候选者。这使得 PrimerStation 能够交互式地搜索和输出合格的引物。 PrimerStation 可以通过寻找更宽的温度范围来选择适合多重 PCR 的高度特异性引物,从而最大限度地减少交叉反应的可能性。它还允许用户在引物设计中添加启发式规则,例如排除引物中的单核苷酸多态性 (SNP),避免 PCR 产物中的 Poly(A) 和 CA 重复,以及使用二级结构预测消除有缺陷的引物。我们进行了多项测试来验证随机选择的 ChrX 引物的 PCR 扩增效果,并确认这些引物完美地扩增了特定的 PCR 产物。
PrimerStation () is a web service that calculates primer sets guaranteeing high specificity against the entire human genome. To achieve high accuracy, we used the hybridization ratio of primers in liquid solution. Calculating the status of sequence hybridization in terms of the stringent hybridization ratio is computationally costly, and no web service checks the entire human genome and returns a highly specific primer set calculated using a precise physicochemical model. To shorten the response time, we precomputed candidates for specific primers using a massively parallel computer with 100 CPUs (SunFire 15 K) about 3 months in advance. This enables PrimerStation to search and output qualified primers interactively. PrimerStation can select highly specific primers suitable for multiplex PCR by seeking a wider temperature range that minimizes the possibility of cross-reaction. It also allows users to add heuristic rules to the primer design, e.g. the exclusion of single nucleotide polymorphisms (SNPs) in primers, the avoidance of poly(A) and CA-repeats in the PCR products, and the elimination of defective primers using the secondary structure prediction. We performed several tests to verify the PCR amplification of randomly selected primers for ChrX, and we confirmed that the primers amplify specific PCR products perfectly.
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发表时间: 2004-03
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影响因子: --
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