PAP-LMPCR for improved, allele-specific footprinting and automated chromatin fine structure analysis.
PAP-LMPCR for improved, allele-specific footprinting and automated chromatin fine structure analysis.
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PAP-LMPCR 用于改进的等位基因特异性足迹和自动化染色质精细结构分析。
DOI：
10.1093/nar/gkm1159
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发表时间：
2008
影响因子：
14.9
通讯作者：
Ingram R
中科院分区：
文献类型：
--
作者：
Ingram R
通过活体印迹和连接介导的pcr （LMPCR）分析染色质精细结构和差异表达基因的转录因子占用是了解染色质对基因表达影响的有力工具。然而，与所有基于pcr的技术一样，由于序列相似性和富含gc或重复序列的存在，实验的准确性经常降低，并且一些序列完全难以分析。在这里，我们描述了一种新的方法，焦磷裂解活化聚合LMPCR或PAP-LMPCR，它能够产生准确和可重复的特异等位基因的足迹，并且可以读取以前无法分析的序列。此外，我们已将该技术用于自动化，从而能够同时快速分析许多不同基因的染色质结构。
The analysis of chromatin fine structure and transcription factor occupancy of differentially expressed genes byin vivofootprinting and ligation-mediated-PCR (LMPCR) is a powerful tool to understand the impact of chromatin on gene expression. However, as with all PCR-based techniques, the accuracy of the experiments has often been reduced by sequence similarities and the presence of GC-rich or repeat sequences, and some sequences are completely refractory to analysis. Here we describe a novel method, pyrophosphorolysis activated polymerization LMPCR or PAP-LMPCR, which is capable of generating accurate and reproducible footprints specific for individual alleles and can read through sequences previously not accessible for analysis. In addition, we have adapted this technique for automation, thus enabling the simultaneous and rapid analysis of chromatin structure at many different genes.