Electroporation and lysis of marine microalga Karenia brevis for RNA extraction and amplification
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), 601-608. (doi:10.1098/rsif.2010.0445). (PMID:21084344).
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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.
|Digital Object Identifier (DOI):||doi:10.1098/rsif.2010.0445|
|Keywords:||electroporation, dielectrophoresis, karenia brevis, rna extraction, rna amplification, dinoflagellate cysts|
|Subjects:||G Geography. Anthropology. Recreation > GC Oceanography
Q Science > QH Natural history > QH301 Biology
T Technology > TK Electrical engineering. Electronics Nuclear engineering
|Divisions:||University Structure - Pre August 2011 > National Oceanography Centre (NERC)
University Structure - Pre August 2011 > School of Electronics and Computer Science
National Oceanography Centre (NERC) > Ocean Technology and Engineering
|Date Deposited:||13 Jan 2011 09:21|
|Last Modified:||27 Mar 2014 19:20|
|RDF:||RDF+N-Triples, RDF+N3, RDF+XML, Browse.|
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