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从感染豚鼠中分离的分枝杆菌的竞争和测序方案。

Protocol for competition and sequencing of mycobacterium isolated from infected guinea pigs.

机构信息

Mycobacterial Signalling Lab, CSIR-Centre for Cellular and Molecular Biology, Habsiguda, Telangana 500007, India; Signal Transduction Lab-I, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, Delhi 110067, India.

Cellular Immunology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, Delhi 110067, India.

出版信息

STAR Protoc. 2022 Oct 29;3(4):101804. doi: 10.1016/j.xpro.2022.101804. eCollection 2022 Dec 16.

DOI:10.1016/j.xpro.2022.101804
PMID:36340884
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9630790/
Abstract

We describe steps for gDNA isolation from mycobacterium strains isolated from guinea pig lungs. We detail steps for infection of guinea pigs with , followed by growth, gDNA isolation, and whole genome sequencing. We also describe an competition experiment to determine the selective advantage of one strain over another. We include details for WGS and mutation spectrum analysis. The protocol can be used to identify mutations that arise in other pathogenic bacteria. For complete details on the use and execution of this protocol, please refer to Naz et al. (2021).

摘要

我们描述了从豚鼠肺部分离的分枝杆菌菌株中提取 gDNA 的步骤。我们详细介绍了用 感染豚鼠，然后进行 生长、gDNA 分离和全基因组测序的步骤。我们还描述了一个 竞争实验，以确定一种菌株相对于另一种菌株的选择性优势。我们包括 WGS 和突变谱分析的详细信息。该方案可用于鉴定其他致病菌中出现的突变。有关此方案的使用和执行的完整详细信息，请参阅 Naz 等人。(2021)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/934ef1ff3406/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/c4ecfe18268d/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/4495add1aba5/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/163c284e5103/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/6577dbe39bdf/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/86e655359810/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/c8e3ca8bbb0f/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/934ef1ff3406/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/c4ecfe18268d/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/4495add1aba5/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/163c284e5103/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/6577dbe39bdf/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/86e655359810/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/c8e3ca8bbb0f/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9b/9630790/934ef1ff3406/gr6.jpg

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