Flow cytometric determination of cell cycle progression via direct labeling of replicated DNA
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F21%3A00538239" target="_blank" >RIV/61388963:_____/21:00538239 - isvavai.cz</a>
Result on the web
<a href="https://doi.org/10.1016/j.ab.2020.114002" target="_blank" >https://doi.org/10.1016/j.ab.2020.114002</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.ab.2020.114002" target="_blank" >10.1016/j.ab.2020.114002</a>
Alternative languages
Result language
angličtina
Original language name
Flow cytometric determination of cell cycle progression via direct labeling of replicated DNA
Original language description
The reported method allows for a simple and rapid monitoring of DNA replication and cell cycle progression in eukaryotic cells in vitro. The DNA of replicating cells is labeled by incorporation of a metabolically-active fluorescent (Cy3) deoxyuridine triphosphate derivative, which is delivered into the cells by a synthetic transporter (SNTT1). The cells are then fixed, stained with DAPI and analyzed by flow cytometry. Thus, this protocol obviates post-labeling steps, which are indispensable in currently used incorporation assays (BrdU, EdU). The applicability of the protocol is demonstrated in analyses of cell cycles of adherent (U-2 OS, HeLa S3, RAW 264.7, J774 A.1, Chem-1, U-87 MG) and suspension (CCRF-CEM, MOLT-4, THP-1, HL-60, JURKAT) cell cultures, including those affected by a DNA polymerase inhibitor (aphidicolin). Owing to a short incorporation time (5–60 min) and reduced number of steps, the protocol can be completed within 1–2 h with a minimal cell loss and with excellent reproducibility.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10401 - Organic chemistry
Result continuities
Project
<a href="/en/project/GA17-14791S" target="_blank" >GA17-14791S: Synthetic analogues of nucleoside triphosphate transporters: development and applications</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2021
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Analytical Biochemistry
ISSN
0003-2697
e-ISSN
1096-0309
Volume of the periodical
614
Issue of the periodical within the volume
Feb 1
Country of publishing house
US - UNITED STATES
Number of pages
9
Pages from-to
114002
UT code for WoS article
000603552000005
EID of the result in the Scopus database
2-s2.0-85097039369