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定量，实时分析，利用分子信标病变特定的细胞裂解液中的碱基切除修复活动

JoVE Journal
Biologia

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JoVE Journal Biologia

Quantitative, Real-time Analysis of Base Excision Repair Activity in Cell Lysates Utilizing Lesion-specific Molecular Beacons

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定量，实时分析，利用分子信标病变特定的细胞裂解液中的碱基切除修复活动

DOI:

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15:01 min

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August 06, 2012

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David Svilar², Conchita Vens, Robert W. Sobol^2,4

¹Department of Pharmacology & Chemical Biology,University of Pittsburgh School of Medicine, ²Hillman Cancer Center,University of Pittsburgh Cancer Institute, ³Department of Experimental Therapy,The Netherlands Cancer Institute, ⁴Department of Human Genetics,University of Pittsburgh School of Public Health

Capitoli

00:05Titolo
02:53Molecular Beacon Design
04:31Nuclear Lysate Preparation
08:31Molecular Beacon Assay
11:51Representative Analysis of Tumor Cell Lysates
13:54Conclusion

Summary

Traduzione automatica

我们描述了DNA糖基化酶和AP内切酶的活动，在核细胞裂解定量，实时测量的方法。该法产生服从动力学分析DNA修复活动率，是适应组织和肿瘤裂解物或纯化蛋白质的DNA修复能力的量化。

Tags

Quantitative Analysis Real-time Measurement Base Excision Repair DNA Glycosylase AP Endonuclease Molecular Beacons Substrate Deoxyoligonucleotide Carboxyfluorescein (6-FAM)Dabcyl Stem-loop Structure Nucleotides Base Pairs Förster Resonance Energy Transfer (FRET)Fluorescence DNA Repair Fluorescence Values Real-time Assessment PCR Instruments

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