Electroporation and lysis of marine microalga Karenia brevis for RNA extraction and amplification
Electroporation and lysis of marine microalga Karenia brevis for RNA extraction and amplification
We describe here a simple device for dielectrophoretic concentration of marine microalga Karenia brevis non-motile cells, followed by electric field-mediated lysis for RNA extraction. The lysate was purified using magnetic beads and pure RNA extracted. RNA quality was assessed off-chip by nucleic acid sequence-based amplification and the optimum conditions for lysis were determined. This procedure will form part of an integrated microfluidic system that is being developed with sub-systems for performing cell concentration and lysis, RNA extraction/purification and real-time quantitative RNA detection. The integrated system and its components could be used for a large range of applications including in situ harmful algal bloom detection, transcriptomics and point-of-care diagnostics.
electroporation, dielectrophoresis, karenia brevis, rna extraction, rna amplification, dinoflagellate cysts
601-608
Bahi, M.M.
97860b79-de5c-443c-b862-45b7bb46bda7
Tsaloglou, M-N.
99ab30ba-15da-4d25-86ba-608d127f8369
Mowlem, M.
6f633ca2-298f-48ee-a025-ce52dd62124f
Morgan, H.
de00d59f-a5a2-48c4-a99a-1d5dd7854174
April 2011
Bahi, M.M.
97860b79-de5c-443c-b862-45b7bb46bda7
Tsaloglou, M-N.
99ab30ba-15da-4d25-86ba-608d127f8369
Mowlem, M.
6f633ca2-298f-48ee-a025-ce52dd62124f
Morgan, H.
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Bahi, M.M., Tsaloglou, M-N., Mowlem, M. and Morgan, H.
(2011)
Electroporation and lysis of marine microalga Karenia brevis for RNA extraction and amplification.
Journal of the Royal Society Interface, 8 (57), .
(doi:10.1098/rsif.2010.0445).
(PMID:21084344)
Abstract
We describe here a simple device for dielectrophoretic concentration of marine microalga Karenia brevis non-motile cells, followed by electric field-mediated lysis for RNA extraction. The lysate was purified using magnetic beads and pure RNA extracted. RNA quality was assessed off-chip by nucleic acid sequence-based amplification and the optimum conditions for lysis were determined. This procedure will form part of an integrated microfluidic system that is being developed with sub-systems for performing cell concentration and lysis, RNA extraction/purification and real-time quantitative RNA detection. The integrated system and its components could be used for a large range of applications including in situ harmful algal bloom detection, transcriptomics and point-of-care diagnostics.
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e-pub ahead of print date: 17 November 2010
Published date: April 2011
Keywords:
electroporation, dielectrophoresis, karenia brevis, rna extraction, rna amplification, dinoflagellate cysts
Organisations:
Ocean Technology and Engineering
Identifiers
Local EPrints ID: 171055
URI: http://eprints.soton.ac.uk/id/eprint/171055
ISSN: 1742-5689
PURE UUID: 045216d4-2e54-401c-a1f3-6897987cb10f
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Date deposited: 13 Jan 2011 09:21
Last modified: 14 Mar 2024 02:48
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Contributors
Author:
M.M. Bahi
Author:
M-N. Tsaloglou
Author:
M. Mowlem
Author:
H. Morgan
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