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在革兰氏负性细菌中生成跨波子插入库，用于高通量测序

JoVE Journal
Biologia

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JoVE Journal Biologia

Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

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在革兰氏负性细菌中生成跨波子插入库，用于高通量测序

DOI:

10.3791/61612-v

•

08:19 min

•

July 07, 2020

•

Misha I. Kazi, Richard D. Schargel, Joseph M. Boll

¹Department of Biology,University of Texas at Arlington

Capítulos

00:04Introduction
00:57Bacterial Mating
02:47Generation of Final Bacterial Mutant Library
04:11DNA Shearing and Poly-C Tail Addition
06:23Results: Transposon Mutant Library
07:31Conclusion

Summary

Tadução automática

我们描述了一种在Gram-阴性细菌中生成饱和变性突变库的方法，以及随后制备用于高通量测序的DNA安培库。例如，我们关注ESKAPE病原体 ，Acineto细菌鲍曼尼，但这个协议是适用于广泛的革兰阴性生物。

Tags

Keywords: Transposon Insertion Library Gram-negative Bacteria High-throughput Sequencing Essential Genes Antibiotic Resistance In Vivo Fitness Mechanical Shearing Poly-CTL Restriction Enzymes Adapter Ligation Bacterial Genetics Genotype-phenotype Relationships Mating Diaminopimelic Acid Glycerol Frozen Stock

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