methBLAST and methPrimerDB: web-tools for PCR based methylation analysis.

methBLAST and methPrimerDB: web-tools for PCR based methylation analysis.
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甲基母细胞和甲基生物症:用于基于PCR的甲基化分析的网络工具。

DOI:
10.1186/1471-2105-7-496
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发表时间:
2006-11-09
期刊:
影响因子:
3
通讯作者:
Vandesompele J
Vandesompele J
中科院分区:
生物学4区
文献类型:
--
作者:
Pattyn F;Hoebeeck J;Robbrecht P;Michels E;De Paepe A;Bottu G;Coornaert D;Herzog R;Speleman F;Vandesompele J

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DNA甲基化通过表观遗传修饰和关键基因的沉默在发育和肿瘤发生中起重要作用。亚硫酸氢盐修饰的DNA上基于PCR的甲基化测定的发展预示着基因甲基化分析在速度和灵敏度上的突破。尽管有这种技术进步,但这些方法需要繁琐的基因引物设计和实验验证。亚硫酸氢盐DNA修饰导致序列改变(所有未甲基化的胞嘧啶转化为尿嘧啶)和一般序列复杂性降低,因为胞嘧啶变得代表性不足。因此,标准BLAST序列同源性搜索不能应用于搜索特异性甲基化引物。为了解决这个问题,我们开发了methBLAST,一个序列相似性搜索程序,基于原始的BLAST算法,但在硅亚硫酸氢盐修饰的基因组序列查询,以评估寡核苷酸序列的相似性。除了引物特异性分析工具,我们还开发了一个名为methPrimerDB的公共数据库,用于存储和检索经验证的基于PCR的甲基化检测。网络界面允许公众免费访问,以执行methBLAST搜索或数据库查询,并提交基于用户的信息。数据库记录可以通过基因符号、核苷酸序列、使用的分析方法、基因或methPrimerDB标识符以及subsequent的名称来搜索。每条记录都包含一个链接,链接到PubMed,以检索有关该基因的其他信息,其基因组背景和描述甲基化测定的文章。为了确保和保持数据的完整性和准确性,该数据库与其他参考数据库相链接。目前,该数据库包含用于研究人类、小鼠和大鼠表观遗传修饰的最流行的基于PCR的甲基化分析方法的引物记录。methPrimerDB和methBLAST可从和获得。我们已经开发了两个集成的和免费提供的基于PCR的甲基化分析的网络工具。methBLAST允许在基于PCR的甲基化测定中对引物特异性进行计算机评估,该甲基化测定可存储在methPrimerDB数据库中,该数据库为经验证的甲基化测定提供了搜索门户。
DNA methylation plays an important role in development and tumorigenesis by epigenetic modification and silencing of critical genes. The development of PCR-based methylation assays on bisulphite modified DNA heralded a breakthrough in speed and sensitivity for gene methylation analysis. Despite this technological advancement, these approaches require a cumbersome gene by gene primer design and experimental validation. Bisulphite DNA modification results in sequence alterations (all unmethylated cytosines are converted into uracils) and a general sequence complexity reduction as cytosines become underrepresented. Consequently, standard BLAST sequence homology searches cannot be applied to search for specific methylation primers. To address this problem we developed methBLAST, a sequence similarity search program, based on the original BLAST algorithm but querying in silico bisulphite modified genome sequences to evaluate oligonucleotide sequence similarities. Apart from the primer specificity analysis tool, we have also developed a public database termed methPrimerDB for the storage and retrieval of validated PCR based methylation assays. The web interface allows free public access to perform methBLAST searches or database queries and to submit user based information. Database records can be searched by gene symbol, nucleotide sequence, analytical method used, Entrez Gene or methPrimerDB identifier, and submitter's name. Each record contains a link to Entrez Gene and PubMed to retrieve additional information on the gene, its genomic context and the article in which the methylation assay was described. To assure and maintain data integrity and accuracy, the database is linked to other reference databases. Currently, the database contains primer records for the most popular PCR-based methylation analysis methods to study human, mouse and rat epigenetic modifications. methPrimerDB and methBLAST are available at and . We have developed two integrated and freely available web-tools for PCR based methylation analysis. methBLAST allows in silico assessment of primer specificity in PCR based methylation assays that can be stored in the methPrimerDB database, which provides a search portal for validated methylation assays.
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发表时间: 2006-01-01
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发表时间: 2005-01-01
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作者:
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