title: Electroporation and lysis of marine microalga Karenia brevis for RNA extraction and amplification.
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.
authors.affiliation: Sensors Development Group, National Oceanography Centre, Southampton, UK. m.tsaloglou@soton.ac.uk
authors.forename: M M
authors.initials: MM
authors.lastname: Bahi
authors.forename: M-N
authors.initials: MN
authors.lastname: Tsaloglou
authors.forename: M
authors.initials: M
authors.lastname: Mowlem
authors.forename: H
authors.initials: H
authors.lastname: Morgan
mesh_terms: D056893:Alveolata / Q000235:genetics*; D018929:Cell Culture Techniques; D018274:Electroporation / Q000379:methods*; D058086:Microalgae / Q000235:genetics*; D044085:Microfluidics / Q000379:methods; D021141:Nucleic Acid Amplification Techniques; D012313:RNA / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D012313:RNA