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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (30): 208-212.doi: 10.11924/j.issn.1000-6850.2012-0979

• 生物技术科学 • 上一篇    下一篇

染色质免疫共沉淀超声波破碎条件的研究

张丽丽 徐碧玉 刘菊华 贾彩红 张建斌 王甲水 金志强   

  • 收稿日期:2012-03-20 修回日期:2012-05-07 出版日期:2012-10-25 发布日期:2012-10-25
  • 基金资助:

    “十二五”农村领域国家科技计划课题;中央级公益性科研院所基本科研业务费专项资金

Exploration of Sonication Conditions during Chromatin Immunoprecipitation

  • Received:2012-03-20 Revised:2012-05-07 Online:2012-10-25 Published:2012-10-25

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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

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 1% 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.