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

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 language	English
Pages (from-to)	50-55
Number of pages	6
Journal	Endocytobiosis and Cell Research
Volume	26
Publication status	Print publication - 2015
Externally published	Yes

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title = "Isolation of macro-nuclear DNA from the rumen ciliate Entodinium caudatum",

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.",

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

year = "2015",

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journal = "Endocytobiosis and Cell Research",

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T1 - Isolation of macro-nuclear DNA from the rumen ciliate Entodinium caudatum

AU - Young, Kenneth

AU - Thomas, Nadine

AU - Duval, Stephan

AU - Newbold, Charles

AU - Fried, Johannes

AU - Brown, David

AU - Hackstein, Johannes

AU - Michalowski, Tadeusz

AU - McEwan, Neil

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

M3 - Article

VL - 26

SP - 50

EP - 55

JO - Endocytobiosis and Cell Research

JF - Endocytobiosis and Cell Research

ER -

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

Abstract

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