Isolation of macro-nuclear DNA from the rumen ciliate Entodinium caudatum

Kenneth Young, Nadine Thomas, Stephan Duval, Charles Newbold, Johannes Fried, David Brown, Johannes Hackstein, Tadeusz Michalowski, Neil McEwan

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Abstract

A method is described which allows isolation of macro‐nuclear DNA from the rumen ciliate Entodinium caudatum. Ciliate cells were enriched from the total microbial population in the rumen of sheep with a single ciliate species present by sedimentation and size filtration. Ciliates were then lysed by homogenisation and the macronuclear component isolated on a Percoll density gradient. Macronuclei can be stored in guanidine. HCl until ready for DNA extraction. DNA which had been extracted from macronuclei fresh from the Percoll gradient, or following guanidine. HCl storage, is of sufficient quality to permit its use for PCR analysis.
Original languageEnglish
Pages (from-to)50-55
Number of pages6
JournalEndocytobiosis and Cell Research
Volume26
Publication statusPrint publication - 2015
Externally publishedYes

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    Cite this

    Young, K., Thomas, N., Duval, S., Newbold, C., Fried, J., Brown, D., Hackstein, J., Michalowski, T., & McEwan, N. (2015). Isolation of macro-nuclear DNA from the rumen ciliate Entodinium caudatum. Endocytobiosis and Cell Research, 26, 50-55.