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| 染色质免疫共沉淀超声波破碎条件的研究 | |
| 引用本文： | 张丽丽,徐碧玉,刘菊华,贾彩红,张建斌,王甲水,金志强.染色质免疫共沉淀超声波破碎条件的研究[J].中国农学通报,2012,28(30):208-212. |
| 作者姓名： | 张丽丽 徐碧玉 刘菊华 贾彩红 张建斌 王甲水 金志强 |
| 作者单位： | 1. 海南大学,海口570228;中国热带农业科学院热带生物技术研究所,农业部热带作物生物学与遗传资源利用重点实验室,海口571101 2. 中国热带农业科学院热带生物技术研究所,农业部热带作物生物学与遗传资源利用重点实验室,海口571101 3. 中国热带农业科学院海口实验站,海口,570102 4. 中国热带农业科学院热带生物技术研究所,农业部热带作物生物学与遗传资源利用重点实验室,海口571101;中国热带农业科学院海口实验站,海口570102 |
| 基金项目： | “十二五”农村领域国家科技计划课题“香蕉分子育种与品种创新”(2011AA10020605);中央级公益性科研院所基本科研业务费专项资金“香蕉转录因子对果实品质形成的调控作用研究”(ITBB110202) |
| 摘 要： | 意在找出拟南芥幼苗适宜的染色质免疫共沉淀(ChIP)超声波破碎的条件，为后期抗体的沉淀提供适当大小的片段。以拟南芥(Columbia ecotype)幼苗为材料，用1%浓度甲醛的缓冲液交联DNA和蛋白质，利用超声波将其染色质随机断裂成适当大小的片段。结果表明，适用于拟南芥幼苗的最佳ChIP试验条件为：用1%甲醛溶液交联DNA和蛋白质的复合物；用65%功率，每次工作20 s，间隔4 min，8次破碎该复合物，可以得到400~800 bp大小的片段，用于后续的试验。经本实验的研究得到了拟南芥幼苗染色质免疫共沉淀超声波破碎的最佳条件，为后续实验奠定了基础。 |
| 关 键 词： | 云南 云南 |
| 收稿时间： | 2012/3/20 0:00:00 |
| 修稿时间： | 5/7/2012 12:00:00 AM |
Exploration of Sonication Conditions during Chromatin Immunoprecipitation |
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| Zhang Lili , Xu Biyu , Liu Juhua , Jia Caihong , Zhang Jianbin , Wang Jiashui , Jin Zhiqiang.Exploration of Sonication Conditions during Chromatin Immunoprecipitation[J].Chinese Agricultural Science Bulletin,2012,28(30):208-212. | |
| Authors: | Zhang Lili Xu Biyu Liu Juhua Jia Caihong Zhang Jianbin Wang Jiashui Jin Zhiqiang |
| Institution: | 2,3(1 Hainan University,Haikou 570228;2 Key Laboratory of Biology and Genetic Resources of Tropical Crops,Ministry of Agriculture;Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101;3 Haikou Experimental Station,Chinese Academy of Tropical Agricultural Sciences,Haikou 570102) |
| Abstract: | This study intended to identify suitable ultrasonic conditions of ChIP which would provide appropriate fragments for the precipitation of antibody in Arabidopsis seedlings （Columbia ecotype）. Arabidopsis thaliana tissues were fixed with lifo concentrations formaldehyde for protein to DNA cross-linking, sonicated to shear the DNA into fragments of appropriate size. The results showed that the optimal sonication conditions for ChIP of Arabidopsis thaliana seedlings turned out to be 8 rounds of 20 seconds each time at 65% power, interval of 4 min each time which was sufficient to obtain 400-800 bp size DNA fragments for the subsequent experiment. Suitable ultrasonic conditions of ChIP were identified through the present study. It laid the foundation for the subsequent experiments. |
| Keywords: | Arabidopsis thaliana chromatin immunoprecipitation sonication conditions formaldehyde crosslinking |
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