Quantitative analysis of deadenylation-independent mRNA decay by a modified MBRACE assay.

Quantitative analysis of deadenylation-independent mRNA decay by a modified MBRACE assay.
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通过改进的 MBRACE 测定对不依赖于去腺苷化的 mRNA 衰减进行定量分析。

DOI:
10.1007/978-1-62703-971-0_28
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发表时间:
2014
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Schoenberg,DanielR
Schoenberg,DanielR
中科院分区:
--
文献类型:
--
作者:
Dougherty,JulieA;Mascarenhas,Roshan;Schoenberg,DanielR

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内切酶裂解是红细胞中含无义人β-珠蛋白mRNA衰变的限速步骤。由此产生的5 '截断的中间产物是聚腺苷化的,比亲本mRNA更稳定。Northern blotting通常用于测量全长mRNA的衰变速率,S1核酸酶保护用于测定衰变中间体的命运。我们采用了更灵敏、更简便的MBRACE测定法(Lasham等人,核酸研究38:e19, 2010),通过检测全长β-珠蛋白及其衰变中间体来定量监测衰变过程。
Endonuclease cleavage is the rate-limiting step in the decay of nonsense-containing human β-globin mRNA in erythroid cells. The 5′-truncated intermediates thus generated are polyadenylated and more stable than the parent mRNA. Northern blotting is commonly used to measure the decay rate of full-length mRNA, and S1 nuclease protection is used to assay the fate of decay intermediates. We have adapted the more sensitive and facile MBRACE assay (Lasham et al., Nucleic Acids Res 38: e19, 2010) to quantitatively monitor the decay process by detecting full-length β-globin and its decay intermediates.
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发表时间: 2007-06-27
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影响因子: 5.1
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