Nuclease P1 digestion/high-performance liquid chromatography, a practical method for DNA quantitation

Journal of Chromatography. a
Olga Shimelis, Roger W Giese

Abstract

We have developed a practical method for quantifying DNA. The method is practical in two ways. First, a single enzyme is used to digest the DNA to nucleotides that are then quantified by HPLC under ordinary conditions. Second, the method quantifies DNA even when it is impure. In our method, "nuclease P1/HPLC," the DNA is hydrolyzed by nuclease P1 and the resulting 2'-deoxynucleoside 5'-monophosphates are quantified by HPLC with UV detection. This method was applied to several kinds of genomic DNA in terms of origin and method by which it had been purified. Calf thymus DNA (purified by salt precipitation by the supplier), pig liver DNA (purified by phenolic extraction or by anion-exchange chromatography using a Genomic Tip from Qiagen) and mouse skin DNA (similarly purified) were tested. In some cases a given sample was purified by two of these methods. The values for the amount of DNA by our method were compared with those by three other methods: acid hydrolysis/HPLC (selected as a reference procedure), UV absorbance, and dye binding. Agreement for all DNA samples between the values by our method versus those provided by acid hydrolysis/HPLC was within 10% for amounts of DNA in the 19-54 microg range. In contrast, UV absorbance...Continue Reading

Citations

Mar 28, 2012·BMC Molecular Biology·Inge Søkilde PedersenPoul Henning Madsen
Sep 6, 2015·Journal of Chromatographic Science·Xiao-li LiLong-jiang Yu
May 7, 2014·Rapid Communications in Mass Spectrometry : RCM·Tanja C W Moerdijk-PoortvlietHenricus T S Boschker
Jun 9, 2009·International Journal of Systematic and Evolutionary Microbiology·Ronald O GarciaRolf Müller
Aug 13, 2014·International Journal of Systematic and Evolutionary Microbiology·Ronald GarciaRolf Müller
Jul 12, 2011·International Journal of Systematic and Evolutionary Microbiology·Kathrin I MohrRolf Müller
Apr 6, 2016·International Journal of Systematic and Evolutionary Microbiology·Ram Prasad AwalRolf Müller
Sep 30, 2016·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Yong MaDonglu Zhang
Dec 3, 2016·International Journal of Systematic and Evolutionary Microbiology·Azam MoradiJoachim Wink
Feb 21, 2018·International Journal of Systematic and Evolutionary Microbiology·Kathrin I MohrJoachim Wink
Jul 21, 2017·Journal of Speech, Language, and Hearing Research : JSLHR·Cynthia R HunterDavid B Pisoni
May 15, 2009·Analytical Chemistry·Julie Farand, Francis Gosselin

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.