PAP-LMPCR: an improved, sequence-selective method for the in vivo analysis of transcription factor occupancy and chromatin fine structure.

PAP-LMPCR: an improved, sequence-selective method for the in vivo analysis of transcription factor occupancy and chromatin fine structure.
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PAP-LMPCR:一种改进的序列选择性方法,用于转录因子占用和染色质精细结构的体内分析。

DOI:
10.1007/978-1-60761-944-4_12
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发表时间:
2011
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
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通讯作者:
Ingram R
Ingram R
中科院分区:
--
文献类型:
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作者:
Ingram R

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体内足迹法和连接介导的PCR(LMPCR)是用于检测真核基因染色质结构的成熟方法。在这里,我们描述了一种改进的方法(焦磷酸解活化聚合LMPCR或PAP-LMPCR),克服了以前的方法的不足,能够通过阅读序列,到目前为止是难治性的这种类型的分析。这包括二核苷酸重复序列或富含GC的区域。我们还描述了能够区分不同等位基因的条件,从而使单等位基因表达的基因的同时分析,而不必采用种间杂交。
In vivo footprinting and ligation-mediated PCR (LMPCR) are well-established methods for the examination of the chromatin structure of eukaryotic genes. Here, we describe an improved method (pyrophosphorolysis activated polymerization LMPCR or PAP-LMPCR) that overcomes the shortfalls of previous methods by being capable of reading through sequences that up to now were refractory to this type of analysis. This includes dinucleotide repeat sequences or GC-rich regions. We also describe conditions capable of distinguishing between different alleles, thus enabling the simultaneous analysis of monoallelically expressed genes without having to employ interspecies hybrids.
通过连接介导的 PCR 测量紫外光产物的形成和修复。
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发表时间: 1999
期刊: Methods in molecular biology (Clifton, N.J.)
影响因子: --
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