id:"swepub:oai:DiVA.org:umu-190404"
Search: id:"swepub:oai:DiVA.org:umu-190404" > Isocratic HPLC anal...
- 1 of 1
- Previous record
- Next record
- To hitlist
Isocratic HPLC analysis for the simultaneous determination of dNTPs, rNTPs and ADP in biological samples
-
- Ranjbarian, Farahnaz (author)
- Umeå universitet,Institutionen för medicinsk kemi och biofysik
-
- Sharma, Sushma (author)
- Umeå universitet,Institutionen för medicinsk kemi och biofysik
-
- Falappa, Giulia (author)
- Umeå universitet,Institutionen för medicinsk kemi och biofysik
- show more...
- show less...
- (creator_code:org_t)
- 2021-11-25
- 2022
- English.
- In: Nucleic Acids Research. - : Oxford University Press. - 0305-1048 .- 1362-4962. ; 50:3
- Journal article (peer-reviewed)
Abstract
Subject headings
Close
- Information about the cellular concentrations of deoxyribonucleoside triphosphates (dNTPs) is instrumental for mechanistic studies of DNA replication and for understanding diseases caused by defects in dNTP metabolism. The dNTPs are measured by methods based on either HPLC or DNA polymerization. An advantage with the HPLC-based techniques is that the parallel analysis of ribonucleoside triphosphates (rNTPs) can serve as an internal quality control of nucleotide integrity and extraction efficiency. We have developed a Freon-free trichloroacetic acid-based method to extract cellular nucleotides and an isocratic reverse phase HPLC-based technique that is able to separate dNTPs, rNTPs and ADP in a single run. The ability to measure the ADP levels improves the control of nucleotide integrity, and the use of an isocratic elution overcomes the shifting baseline problems in previously developed gradient-based reversed phase protocols for simultaneously measuring dNTPs and rNTPs. An optional DNA-polymerase-dependent step is used for confirmation that the dNTP peaks do not overlap with other components of the extracts, further increasing the reliability of the analysis. The method is compatible with a wide range of biological samples and has a sensitivity better than other UV-based HPLC protocols, closely matching that of mass spectrometry-based detection.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
Find in a library
- Nucleic Acids Research (Search for host publication in LIBRIS)
To the university's database
- 1 of 1
- Previous record
- Next record
- To hitlist
Find more in SwePub
- By the author/editor
- Ranjbarian, Fara ...
- Sharma, Sushma
- Falappa, Giulia
- Taruschio, Walte ...
- Chabes, Andrei, ...
- Hofer, Anders
- About the subject
-
- MEDICAL AND HEALTH SCIENCES
- MEDICAL AND HEAL ...
- and Basic Medicine
- and Cell and Molecul ...
-
- NATURAL SCIENCES
- NATURAL SCIENCES
- and Biological Scien ...
- and Biochemistry and ...
- Articles in the publication
- Nucleic Acids Re ...
- By the university
- Umeå University
Search outside SwePub
- Extend your search to:
- Google Book Search
- Google Scholar
Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.
- Copyright © LIBRIS - National Library Systems
- LIBRIS.kb.se
