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