Nuclei isolation protocols for flow cytometry allowing nuclear DNA content estimation in problematic microalgal groups

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F21%3A10440406" target="_blank" >RIV/00216208:11310/21:10440406 - isvavai.cz</a>

Result on the web

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=qTzs7C3yRb" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=qTzs7C3yRb</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s10811-021-02433-z" target="_blank" >10.1007/s10811-021-02433-z</a>

Result language

angličtina

Original language name

Nuclei isolation protocols for flow cytometry allowing nuclear DNA content estimation in problematic microalgal groups

Original language description

Microalgae are fundamentally important organisms for global ecosystem functioning with high potential in biotechnology and its applications. The knowledge of their nuclear DNA content has become a prerequisite for many areas of microalgal research. Due to common presence of various pigments, secondary metabolites and complex cell walls, the nuclear DNA content estimation using flow cytometry (FCM) is, however, often laborious or even impossible with the currently used protocols. In this study the performance of six nuclei isolation protocols was compared on various problematic microalgae using FCM. The nuclei isolation methods involved osmotic bursting of cells, razor blade chopping of fresh biomass and two newly introduced protocols, razor blade chopping of desiccated biomass and bead beating. These techniques also involved the use of two different nuclei isolation solutions, Otto I + II solutions, and LB01 buffer. Performance of the particular protocols differed greatly, depending on the used nuclei isolation solution and microalgal group. The most successful method was a newly adopted chopping of desiccated biomass in LB01 buffer. This method seems more appropriate for nuclei isolation in filamentous microalgae; on the other hand, bead beating appears to be more suitable for nuclei isolation in solitarily living algae. Using the optimal protocol for a given species, their nuclear DNA content was estimated, resulting in first DNA content estimates for four investigated taxa (Chlamydomonas noctigama, Gonyostomum semen, Microglena sp. and Stigeoclonium sp.). The estimated DNA content spanned from 0.15 to 32.52 pg.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10611 - Plant sciences, botany

Project

—

Continuities

S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2021

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Applied Phycology

ISSN

0921-8971

e-ISSN

—

Volume of the periodical

33

Issue of the periodical within the volume

4

Country of publishing house

NL - THE KINGDOM OF THE NETHERLANDS

Number of pages

11

Pages from-to

2057-2067

UT code for WoS article

000628087700004

EID of the result in the Scopus database

2-s2.0-85102552851