Abstract
Bacterial artificial chromosome (BAC) library is an important genomic resource useful in targeted marker development, positional cloning, physical mapping and a substrate for genome sequencing for better understanding the genome organization of a species. The present manuscript elucidates the improvement in protocols for economical and efficient BAC insert DNA isolation, BAC end sequencing and FISH for physical localization on the metaphase chromosome complements. BAC clones of Clarias magur, maintained in 384—well plate format in our laboratory, were used in this study. The protocols gave consistent and efficient results. We use routinely these protocols for BAC insert DNA extraction, generating end sequence data of the clone and constructing DNA probes to hybridize on the metaphase spreads of C. magur using FISH for physical their localization.
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Acknowledgements
Authors are highly thankful to the Director, ICAR-National Bureau of Fish Genetic Resources, Lucknow, for infrastructure support and encouragement for taking up the work. The authors are also thankful to the CABin Scheme of ICAR, implemented through CABin Division of ICAR-IASRI, New Delhi, for financial support of the project in which the work was undertaken.
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Kumar, R., Baisvar, V.S., Kushwaha, B. et al. Improved protocols for BAC insert DNA isolation, BAC end sequencing and FISH for construction of BAC based physical map of genes on the chromosomes. Mol Biol Rep 47, 2405–2413 (2020). https://doi.org/10.1007/s11033-020-05283-z
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DOI: https://doi.org/10.1007/s11033-020-05283-z