Abstract
Macroarray-based analysis is a powerful and economic format to study variations in “clustered regularly interspaced short palindromic repeat (CRISPR)” loci in bacteria. To date, it was used almost exclusively for Mycobacterium tuberculosis and was named spoligotyping (spacer oligonucleotides typing). Here, we describe the pipeline of this approach that includes search of loci and selection of spacers, preparation of the membrane with immobilized probes and spoligotyping itself (PCR and reverse hybridization).
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Acknowledgements
We are grateful to Thierry Zozio, Elisabeth Streit and Julie Millet (Institut Pasteur de la Guadeloupe), Anna Vyazovaya and Olga Narvskaya (St. Petersburg Pasteur Institute) for helpful discussions. Igor Mokrousov gratefully acknowledges support from Russian Science Foundation (project 14-14-00292).
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Mokrousov, I., Rastogi, N. (2015). Spacer-Based Macroarrays for CRISPR Genotyping. In: Lundgren, M., Charpentier, E., Fineran, P. (eds) CRISPR. Methods in Molecular Biology, vol 1311. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2687-9_7
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DOI: https://doi.org/10.1007/978-1-4939-2687-9_7
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