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酵母中转录因子结合位点的鉴定:高密度寡核苷酸和基于PCR的微阵列平台的比较

Transcription factor binding site identification in yeast: a comparison of high-density oligonucleotide and PCR-based microarray platforms.

作者信息

Borneman Anthony R, Zhang Zhengdong D, Rozowsky Joel, Seringhaus Michael R, Gerstein Mark, Snyder Michael

机构信息

Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06511, USA.

出版信息

Funct Integr Genomics. 2007 Oct;7(4):335-45. doi: 10.1007/s10142-007-0054-7. Epub 2007 Jul 19.

Abstract

In recent years, techniques have been developed to map transcription factor binding sites using chromatin immunoprecipitation combined with DNA microarrays (chIP chip). Initially, polymerase chain reaction (PCR)-based DNA arrays were used for the chIP chip procedure, however, high-density oligonucleotide (HDO) arrays, which allow for the production of thousands more features per array, have emerged as a competing array platform. To compare the two platforms, data from chIP chip analysis performed for three factors (Tec1, Ste12, and Sok2) using both HDO and PCR arrays under identical experimental conditions were compared. HDO arrays provided increased reproducibility and sensitivity, detecting approximately three times more binding events than the PCR arrays while also showing increased accuracy. The increased resolution provided by the HDO arrays also allowed for the identification of multiple binding peaks in close proximity and of novel binding events such as binding within ORFs. The HDO array platform provides a far more robust array system by all measures than PCR-based arrays, all of which is directly attributable to the large number of probes available.

摘要

近年来,已开发出利用染色质免疫沉淀结合DNA微阵列(芯片)来绘制转录因子结合位点的技术。最初,基于聚合酶链反应(PCR)的DNA阵列用于芯片程序,然而,高密度寡核苷酸(HDO)阵列已成为一种具有竞争力的阵列平台,它每个阵列能产生数千个更多的特征。为了比较这两种平台,对在相同实验条件下使用HDO和PCR阵列针对三个因子(Tec1、Ste12和Sok2)进行的芯片分析数据进行了比较。HDO阵列提供了更高的重现性和灵敏度,检测到的结合事件比PCR阵列多大约三倍,同时还显示出更高的准确性。HDO阵列提供的更高分辨率还允许识别紧密相邻的多个结合峰以及新的结合事件,如开放阅读框内的结合。从各方面衡量,HDO阵列平台都提供了一个比基于PCR的阵列强大得多的阵列系统,所有这些都直接归因于可用探针的数量众多。

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