Colony hybridization

Hybridization is applied to the nucleic acid released from microbial colonies and labelled with a probe for detection by methods such as ultraviolet light or autoradiography. This is great for screening clones.
The process of colony hybridization: growth of cell colonies, replication on filter, hybridization, and identification of desired colonies.

Colony hybridization is a method of selecting bacterial colonies with desired genes.[1] This method was discovered by Michael Grunstein and David S. Hogness. [2]

Method

Colony hybridization begins with culturing sparsely populated bacterial colonies on a nutrient agar plate. These colonies are symmetrically replicated on a nitrocellulose filter by direct contact, after which the cells on the filter membrane are lysed and their DNA is denatured, allowing it to bind to the filter. These DNA clusters are then hybridized to a desired radioactively-labelled RNA or DNA probe (chosen specifically beforehand) and screened by autoradiography. DNA clusters that exhibit a desired gene are then matched up to the corresponding (living) bacterial colonies, which can be isolated for further growth and experimentation.[2]

References

  1. ↑ Rédei, George P. (16 July 2016). Encyclopedia of Genetics, Genomics, Proteomics and Informatics. Springer Netherlands. p. 392. doi:10.1007/978-1-4020-6754-9. ISBN 978-1-4020-6754-9.
  2. 1 2 Grunstein, M; Hogness, D S (1975). "Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene". Proceedings of the National Academy of Sciences. 72 (10): 3961–3965. Bibcode:1975PNAS...72.3961G. doi:10.1073/pnas.72.10.3961. PMC 433117. PMID 1105573.


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