RNA Extraction from Soil Bacterium Pseudomonas Putida And Green Alga Raphidocelis Subcapitata After Exposure to Nanoscale Zero Valent Iron

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F46747885%3A24620%2F21%3A00009748" target="_blank" >RIV/46747885:24620/21:00009748 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.confer.cz/nanocon/2020/3764-rna-extraction-from-soil-bacterium-pseudomonas-putida-and-green-alga-raphidocelis-subcapitata-after-exposure-to-nzvi" target="_blank" >https://www.confer.cz/nanocon/2020/3764-rna-extraction-from-soil-bacterium-pseudomonas-putida-and-green-alga-raphidocelis-subcapitata-after-exposure-to-nzvi</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.37904/nanocon.2020.3764" target="_blank" >10.37904/nanocon.2020.3764</a>

Jazyk výsledku

angličtina

Název v původním jazyce

RNA Extraction from Soil Bacterium Pseudomonas Putida And Green Alga Raphidocelis Subcapitata After Exposure to Nanoscale Zero Valent Iron

Popis výsledku v původním jazyce

Soil bacterium Pseudomonas putida and green alga Raphidocelis subcapitata are often used in ecotoxicology studies, including assessment of nanomaterials. Exposure of these microorganisms to reactive nZVI, however, reduce the quality of extracted RNA for further transcriptomic analysis. In this study, we compare RNA extraction using eight extraction protocols. The extraction using the commercial column kits was often unable to elute both the RNA species in a single elution, whereas phenol-chloroform method using RNAzol recovered both small RNA and large RNA from P. putida. On the other hand, the column based RNA extraction kit from ISOLATE II RNA Mini gave the best yield approximately of 1 µg/µl of RNA and RIN 9.0. RNA extraction from R. subcapitata was more complicated when we used a similar column based RNA extraction system, it required additional and thorough cell lysis steps that included cell lyophilisation and ribolization prior to phenol-chloroform extraction method. Then the yield of an effective RNA concentration ranged from 300 ng/µl to 980 ng/µl, giving the RIN 5.5-7.8. The final protocols solved most of the extraction problems, and allow experiments involving RNA gene expression analysis of microorganisms exposed to reactive nZVI.

Název v anglickém jazyce

RNA Extraction from Soil Bacterium Pseudomonas Putida And Green Alga Raphidocelis Subcapitata After Exposure to Nanoscale Zero Valent Iron

Popis výsledku anglicky

Soil bacterium Pseudomonas putida and green alga Raphidocelis subcapitata are often used in ecotoxicology studies, including assessment of nanomaterials. Exposure of these microorganisms to reactive nZVI, however, reduce the quality of extracted RNA for further transcriptomic analysis. In this study, we compare RNA extraction using eight extraction protocols. The extraction using the commercial column kits was often unable to elute both the RNA species in a single elution, whereas phenol-chloroform method using RNAzol recovered both small RNA and large RNA from P. putida. On the other hand, the column based RNA extraction kit from ISOLATE II RNA Mini gave the best yield approximately of 1 µg/µl of RNA and RIN 9.0. RNA extraction from R. subcapitata was more complicated when we used a similar column based RNA extraction system, it required additional and thorough cell lysis steps that included cell lyophilisation and ribolization prior to phenol-chloroform extraction method. Then the yield of an effective RNA concentration ranged from 300 ng/µl to 980 ng/µl, giving the RIN 5.5-7.8. The final protocols solved most of the extraction problems, and allow experiments involving RNA gene expression analysis of microorganisms exposed to reactive nZVI.

Druh

D - Stať ve sborníku

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/LM2018124" target="_blank" >LM2018124: Nanomateriály a nanotechnologie pro ochranu životního prostředí a udržitelnou budoucnost</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2021

Kód důvěrnosti údajů

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

Název statě ve sborníku

12TH INTERNATIONAL CONFERENCE ON NANOMATERIALS - RESEARCH & APPLICATION (NANOCON 2020)

ISBN

978-80-87294-98-7

ISSN

2694-930X

e-ISSN

—

Počet stran výsledku

6

Strana od-do

477-482

Název nakladatele

TANGER LTD

Místo vydání

—

Místo konání akce

Brno, Czech Republic

Datum konání akce

1. 1. 2020

Typ akce podle státní příslušnosti

WRD - Celosvětová akce

Kód UT WoS článku

000664505500081