| 鸡脂肪组织总RNA提取方法的优化和RT-PCR检测 |
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| 引用本文： | 邢晋祎,姜运良.鸡脂肪组织总RNA提取方法的优化和RT-PCR检测[J].吉林农业科学,2009,34(5):41-44. |
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| 作者姓名： | 邢晋祎 姜运良 |
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| 作者单位： | 1. 临沂师范学院生命科学学院,山东,临沂,276005
2. 山东农业大学动物科技学院,山东,泰安,271018 |
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| 基金项目： | 临沂师范学院博士启动基金 |
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| 摘 要： | 提取完整的RNA是基因表达分析中的基础,但由于脂肪组织含有大量的油脂,所以从脂肪组织中提取高质量的RNA有一定的困难。在本研究中,报道了经过优化TRIzol试剂操作程序成功地从肉鸡脂肪中分离出了完整、高质量、纯度好的总RNA。用该方法提取的RNA显示清晰的28S、18S和5S三条带,OD260/OD280均值为1.893,浓度均值为263μg/mL。保温试验与RT-PCR扩增鸡LPL和GAPDH基因的分析结果也证明了所提取的总RNA质量较高。
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| 关 键 词： | 鸡 脂肪 RNA 优化 |
Optimization of Methods for Total RNA Extraction from Chicken Adipose and RT-PCR Detection |
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| XING Jin-yi,JIANG Yun-liang.Optimization of Methods for Total RNA Extraction from Chicken Adipose and RT-PCR Detection[J].Jilin Agricultural Sciences,2009,34(5):41-44. |
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| Authors: | XING Jin-yi JIANG Yun-liang |
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| Abstract: | Extraction of intact RNA is essential for gene expression analysis. The isolation of high-quality RNA from adipose tissue is difficult due to numerous lipids. In this paper, we reported a protocol which we have optimized based on the Invitrogen TRIzol procedures to produce high yields of purified fully intact RNA from broilers abdominal adipose. The total RNA isolated using this procedure demonstrated sharp, undegraded 28S, 18S, and 5S ribosomal RNA bands. The average ratio OD260/OD280 and concentration of RNA were 1.893 and 263 μg/mL, respectively. Successful RNA extraction was also observed by heat preservation experiment. RT-PCR analysis of chicken LPL and GAPDH genes was used to verify the quality of the RNA. |
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| Keywords: | RNA |
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