Simple, fast, cost-efficient, reliable, and highly automated DNA content analysis of cells in adherent cultures

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F24%3A00585791" target="_blank" >RIV/61388963:_____/24:00585791 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/75010330:_____/24:00014696 RIV/60162694:G44__/25:00563332 RIV/00216208:11110/24:10482396 RIV/00064211:_____/24:W0000011

Výsledek na webu

<a href="https://doi.org/10.1002/cyto.a.24840" target="_blank" >https://doi.org/10.1002/cyto.a.24840</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/cyto.a.24840" target="_blank" >10.1002/cyto.a.24840</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Simple, fast, cost-efficient, reliable, and highly automated DNA content analysis of cells in adherent cultures

Popis výsledku v původním jazyce

The most commonly used flow cytometric (FCM) analysis of cellular DNA content relies on ethanol fixation followed by RNA digestion and propidium iodide (PI) intercalation into double-stranded DNA. This is a laborious and time-consuming procedure that is subject to systematic errors due to centrifugation and washing steps associated with sample preparation. It can adversely affect the reliability of the results. Here, we present a modified concept of DNA quantification in adherent cell lines by FCM that involves neither ethanol fixation nor any washing and cell transferring steps. Our high throughput assay of adherent cell lines reduces sample-processing time, requires minimal workload, provides a possibility for automation, and, if needed, also allows a significant reduction in the size of individual samples. Working with a well-proven commercial tool-The BD Cycletest Plus DNA Reagent Kit-primarily designed for cell cycle analysis and aneuploidy determination in experimental and clinical samples, we suggest a novel, very efficient, and robust approach for DNA research in adherent cell cultures.

Název v anglickém jazyce

Simple, fast, cost-efficient, reliable, and highly automated DNA content analysis of cells in adherent cultures

Popis výsledku anglicky

The most commonly used flow cytometric (FCM) analysis of cellular DNA content relies on ethanol fixation followed by RNA digestion and propidium iodide (PI) intercalation into double-stranded DNA. This is a laborious and time-consuming procedure that is subject to systematic errors due to centrifugation and washing steps associated with sample preparation. It can adversely affect the reliability of the results. Here, we present a modified concept of DNA quantification in adherent cell lines by FCM that involves neither ethanol fixation nor any washing and cell transferring steps. Our high throughput assay of adherent cell lines reduces sample-processing time, requires minimal workload, provides a possibility for automation, and, if needed, also allows a significant reduction in the size of individual samples. Working with a well-proven commercial tool-The BD Cycletest Plus DNA Reagent Kit-primarily designed for cell cycle analysis and aneuploidy determination in experimental and clinical samples, we suggest a novel, very efficient, and robust approach for DNA research in adherent cell cultures.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2024

Kód důvěrnosti údajů

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

Název periodika

Cytometry. Part A

ISSN

1552-4922

e-ISSN

1552-4930

Svazek periodika

105

Číslo periodika v rámci svazku

6

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

6

Strana od-do

474-479

Kód UT WoS článku

001217258600001

EID výsledku v databázi Scopus

2-s2.0-85192204537