Maps of histone variant H3.V deposition in Trypanosoma brucei brucei. Maps of histone variant H3.V deposition in Trypanosoma brucei brucei

Chromatin immunoprecipitation of genomic loci in Trypanosoma brucei where histone variant H3.V is deposited. This was achieved by deletion of one H3.V allele and N-terminal tagging of the second H3.V allele with a Ty1 tag. During the ChIP experiment, the DNA was digested with MNase to obtain mononucleosomes. Nucleosomes containing H3.V were pulled down by using a BB2 anti-Ty1 antibody. Cross links are reversed and mononucleosomal DNA is purified and prepared for Illumina sequencing. Overall design: Mnase ChIPs were done in duplicates, input control samples were included for each ChIP sample.

针对布氏锥虫（Trypanosoma brucei）内组蛋白变体H3.V沉积的基因组位点开展染色质免疫共沉淀（Chromatin immunoprecipitation, ChIP）实验。本实验通过敲除一个H3.V等位基因，并以Ty1标签对另一拷贝的H3.V等位基因进行N端标记构建实验样本。在ChIP实验流程中，使用微球菌核酸酶（Micrococcal nuclease, MNase）酶切DNA以获取单核小体；通过BB2抗Ty1抗体下拉富集携带H3.V的核小体。随后逆转交联反应，纯化单核小体DNA并制备用于Illumina测序的文库。实验整体设计：MNase ChIP实验设置生物学重复，且每个ChIP样本均配套设置Input对照样本。