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线性扩增介导的PCR - 未知侧翼DNA的遗传物质和表征本地化

JoVE Journal
Biologia

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JoVE Journal Biologia

Linear Amplification Mediated PCR – Localization of Genetic Elements and Characterization of Unknown Flanking DNA

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线性扩增介导的PCR - 未知侧翼DNA的遗传物质和表征本地化

DOI:

10.3791/51543-v

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11:58 min

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June 25, 2014

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Richard Gabriel, Ina Kutschera, Cynthia C Bartholomae, Christof von Kalle, Manfred Schmidt

¹Department of Translational Oncology,National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ)

Capitoli

00:05Titolo
01:33Preamplifying the Vector Genome Junctions
03:19Magnetic Separation of the PCR Products
04:39LAM PCR Preparation
07:39Non-restrictive LAM PCR Preparation
08:38Exponential Amplifications and Product Clean-Up
10:41Electrophoresis Analysis and Vector Insertion Site Analysis
11:26Conclusion

Summary

Traduzione automatica

线性扩增介导的（LAM）-PCR方法是开发，以确定基因组中的整合型病毒载体的精确位置的方法。该技术已经发展成为学习中的基因治疗的患者中，新颖的矢量技术，T-细胞的多样性，癌症干细胞模型等生物安全克隆动力学的优越方法

Tags

Linear Amplification Mediated PCR LAM-PCR Integration Site Analysis Gene Therapy Hematopoiesis Clonal Fate Proto-oncogene Retroviral Vector Lentiviral Vector Genome Integration Unknown Flanking DNA

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