跳转至内容
Merck
CN

Histone Modification ChIP-seq on Arabidopsis thaliana Plantlets.

Bio-protocol (2021-12-04)
Juan S Ramirez-Prado, David Latrasse, Moussa Benhamed
摘要

Characterizing the molecular mechanisms regulating gene expression is crucial for understanding the regulatory processes underlying physiological responses to environmental and developmental signals in eukaryotes. The covalent modification of histones contributes to the compaction levels of chromatin, as well as the recruitment of the transcriptional machinery to specific loci, facilitating metastable changes in gene activity. ChIP-seq (Chromatin Immunoprecipitation followed by sequencing) has become the gold standard method for determining histone modification profiles among different organisms, tissues, and genotypes. In the current protocol, we describe a highly robust method for performing ChIP-seq of histone modifications in Arabidopsis thaliana plantlets. Besides its robustness, this method uses in-house-prepared buffers for chromatin extraction, immunoprecipitation, washing, and elusion, making it cost-effective in contrast to commercial kits.

材料
产品编号
品牌
产品描述

Roche
不含EDTA的cOmplete蛋白酶抑制剂混合物, Tablets provided in glass vials
Sigma-Aldrich
乙二胺四乙酸 二钠盐 溶液, BioUltra, pH 8.0, ~0.5 M in H2O
Sigma-Aldrich
抗-三甲基-组蛋白H3 (Lys27)抗体, Upstate®, from rabbit
Sigma-Aldrich
甘氨酸, ACS reagent, ≥98.5%
Sigma-Aldrich
抗三甲基组蛋白H3(Lys4)抗体, Upstate®, from rabbit
Sigma-Aldrich
抗乙酰组蛋白H3(Lys14)抗体, serum, Upstate®
Sigma-Aldrich
抗乙酰组蛋白H3(Lys9)抗体, serum, Upstate®
Sigma-Aldrich
抗-单甲基-组蛋白H3(Lys27)抗体, Upstate®, from rabbit