Inicio


Publicación

Immobilization of microalgae cells in alginate facilitates isolation of DNA and RNA


Información de la publicación

Información de la publicación
Tipo de publicación

Científica

Tipología

Investigación y estudios

Medio de publicación

Impreso: Artículos de investigación científica o tecnológica T1

Resumen

Isolation of nucleic acids fromChlorella is difficult, given the chemically complex nature of their cellwalls and variable production of metabolites. Immobilization of microalgae in polymers adds additional difficulty. Here, we modified, amended, and standardized methods for isolation of nucleic acids and compared the yield of DNA and RNA fromfree-living and encapsulated microalgae C. sorokiniana. Isolation of nucleic acids fromimmobilized cells required twosteps in dissolving the alginatematrix, releasing the cells, andmechanical disruption with glass beads. For DNA extraction, we used modified versions of a commercial kit along with the hexadecyltrimethylammonium bromide (CTAB) method. For RNA extraction, we used the commercial TRI reagent procedure and the CTAB-dithiotreitol method. Quantity and quality of nucleic acids in extracts varied with growth conditions, isolation procedures, and time of incubation of the original  culture. There were consistently higher amounts of DNA and RNA in extracts from immobilized cells. Quantitatively, the modified procedure with the commercial Promega kit was the most reliable procedure for isolating DNA and a modified commercial TRI reagent procedure was the choice for isolating RNA. All four procedures eliminated proteins efficiently and had lowlevels of contamination fromresidual polysaccharides fromthematrices and/ormetabolites naturally produced by the microalgae. All DNA extracts under both growth conditions, time of incubation, and two isolation methods successfully amplified the 18S ribosomal RNA by PCR and quantitative reverse transcription (RT-qPCR).

Autores

Blanca R. Lopez
Juan-Pablo Hernandez
Yoav Bashan
Luz E. de-Bashan

Registro ISSN

0167-7012

DOI Artículo Digital Immobilization of microalgae cells in alg...NA and RNA
Isolation of nucleic acids fromChlorella is difficult, given the chemically complex nature of their cellwalls and variable production of metabolites. Immobilization of microalgae in polymers adds additional difficulty. Here, we modified, amended, and stan
SNIES Área

Environmental Science

SNIES Categoría

Molecular Biology

Fecha de publicación 21 de febrero de 2017
Fecha de aceptación 10 de febrero de 2017
Medio indexado (nombre)

Journal of Microbiological Methods

Bases de datos donde está referenciada

sciencedirect

English information
Title

Immobilization of microalgae cells in alginate facilitates isolation of DNA and RNA

Abstract

Isolation of nucleic acids fromChlorella is difficult, given the chemically complex nature of their cellwalls and variable
production of metabolites. Immobilization of microalgae in polymers adds additional difficulty. Here, we
modified, amended, and standardized methods for isolation of nucleic acids and compared the yield of DNA
and RNA fromfree-living and encapsulated microalgae C. sorokiniana. Isolation of nucleic acids fromimmobilized
cells required twosteps in dissolving the alginatematrix, releasing the cells, andmechanical disruption with glass
beads. For DNA extraction, we used modified versions of a commercial kit along with the
hexadecyltrimethylammonium bromide (CTAB) method. For RNA extraction, we used the commercial TRI reagent
procedure and the CTAB-dithiotreitol method. Quantity and quality of nucleic acids in extracts varied
with growth conditions, isolation procedures, and time of incubation of the original culture. There were consistently
higher amounts of DNA and RNA in extracts from immobilized cells. Quantitatively, the modified procedure
with the commercial Promega kit was the most reliable procedure for isolating DNA and a modified
commercial TRI reagent procedure was the choice for isolating RNA. All four procedures eliminated proteins efficiently
and had lowlevels of contamination fromresidual polysaccharides fromthematrices and/ormetabolites
naturally produced by the microalgae. All DNA extracts under both growth conditions, time of incubation, and
two isolation methods successfully amplified the 18S ribosomal RNA by PCR and quantitative reverse transcription
(RT-qPCR).

Keywords

Chlorella, Immobilization, Microalgae, Nucleic acid

Información de contacto

Contacto de Publicaciones