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

Young, Kenneth W.; Thomas, Nadine A.; Duval, Stephane M.; Newbold, C. Jamie; Fried, Johannes; Brown, David S.; Hackstein, Johannes H.P.; Michalowski, Tadeusz; McEwan, Neil R.

Authors

Kenneth W. Young

Nadine A. Thomas

Stephane M. Duval

C. Jamie Newbold

Johannes Fried

David S. Brown

Johannes H.P. Hackstein

Tadeusz Michalowski

Neil R. McEwan



Abstract

A method is described which allows isolation of macronuclear 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.

Citation

YOUNG, K.W., THOMAS, N.A., DUVAL, S.M., NEWBOLD, J., FRIED, J., BROWN, D.S., HACKSTEIN, J.H.P., MICHALOWSKI, T. and MCEWAN, N.R. 2015. Isolation of macro-nuclear DNA from the rumen ciliate Entodinium caudatum. Endocytobiosis and cell research [online], 26, pages 50-55. Available from: https://zs.thulb.uni-jena.de/receive/jportal_jparticle_00355360

Journal Article Type Article
Acceptance Date Jun 15, 2015
Online Publication Date Dec 31, 2015
Publication Date Dec 31, 2015
Deposit Date Dec 14, 2015
Publicly Available Date Dec 14, 2015
Journal Endocytobiosis and cell research
Print ISSN 0256-1514
Electronic ISSN 1613-8872
Publisher International Society of Endocytobiology
Peer Reviewed Peer Reviewed
Volume 26
Pages 50-55
Keywords Macronucleus; Ciliate; Entodinium caudatum
Public URL http://hdl.handle.net/10059/3256
Publisher URL https://zs.thulb.uni-jena.de/receive/jportal_jparticle_00355360

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