Determination of 5-methylcytosine by acid hydrolysis of DNA with hydrofluoric acid

Catania, Jason; Keenan, B C; Margison, Geoffrey P.; Fairweather, D.S.

doi:10.1016/0003-2697(87)90175-8

Cited by 27 publications

(16 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…LC-ESI-MS/MS needs less DNA than the other methods (1 µg for LC-ESI-MS/MS vs 5–10 µg for HPLC) and requires less time per sample (15–60 min for separation using HPLC vs <15 min for separation using LC-ESI-MS/MS). 23 , 24 Direct measurement of 5-methycytosine content in DNA requires a large amount of DNA and is labor intensive, which led to the development of an indirect method that measures methylation levels of repetitive elements ( ALU , LINE1 , and SAT ). The repetitive elements represent over 45% of CpG dinucleotides in the human genome and are correlated with 5-methylcytosine levels throughout the genome.…”

Section: Issues In the Development Of Methylation-based Biomarkersmentioning

confidence: 99%

DNA Methylation in Peripheral Blood: A Potential Biomarker for Cancer Molecular Epidemiology

Lian

Choi

Lee

et al. 2012

Journal of Epidemiology

132

104

View full text Add to dashboard Cite

Aberrant DNA methylation is associated with cancer development and progression. There are several types of specimens from which DNA methylation pattern can be measured and evaluated as an indicator of disease status (from normal biological process to pathologic condition) and even of pharmacologic response to therapy. Blood-based specimens such as cell-free circulating nucleic acid and DNA extracted from leukocytes in peripheral blood may be a potential source of noninvasive cancer biomarkers. In this article, we describe the characteristics of blood-based DNA methylation from different biological sources, detection methods, and the factors affecting DNA methylation. We provide a comprehensive literature review of blood-based DNA methylation as a cancer biomarker and focus on the study of DNA methylation using peripheral blood leukocytes. Although DNA methylation patterns measured in peripheral blood have great potential to be useful and informative biomarkers of cancer risk and prognosis, large systematic and unbiased prospective studies that consider biological plausibility and data analysis issues will be needed in order to develop a clinically feasible blood-based assay.

show abstract

Section: Issues In the Development Of Methylation-based Biomarkersmentioning

confidence: 99%

DNA Methylation in Peripheral Blood: A Potential Biomarker for Cancer Molecular Epidemiology

Lian

Choi

Lee

et al. 2012

Journal of Epidemiology

132

104

View full text Add to dashboard Cite

show abstract

“…(11,12). The third approach is to hydrolyze DNA enzymatically, resulting in nucleotides or nucleosides (i.e., methyl-deoxycitidine) (10,(13)(14)(15)(16)(17)(18)(19)(20) or by acid hydrolysis (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30), resulting in the free bases (i.e., cytosine, the only base that can be methylated to form 5-methylcytosine).…”

Section: Introductionmentioning

confidence: 99%

“…Detection and quantification of the free bases has been performed with capillary electrophoresis (21), high-resolution MS (22), gas chromatography (GC) (23,24), HPLC (28)(29)(30) and GC-MS (25-27, 31, 32).…”

Section: Introductionmentioning

confidence: 99%

Global DNA methylation measured by liquid chromatography-tandem mass spectrometry: analytical technique, reference values and determinants in healthy subjects

Kok¹,

Smith²,

Barto³

et al. 2007

Clinical Chemical Laboratory Medicine

View full text Add to dashboard Cite

show abstract

“…[ [33][34][35][36] The latter is not a preferred option due to the potential of contamination from 5-methylcytosine derived from RNA that maybe present in the DNA sample which is also released by acid hydrolysis.…”

Section: Introductionmentioning

confidence: 99%

Determination of 5-methyl-2′-deoxycytidine in genomic DNA using high performance liquid chromatography-ultraviolet detection

Sandhu

Kaur

Armstrong

et al. 2009

Journal of Chromatography B

View full text Add to dashboard Cite

2 Abbreviations: 5-MedC, 5-methyl-2′-deoxycytidine; dC, 2′-deoxycytidine; dG, 2′-deoxyguanosine; T, thymidine; dA, 2′-deoxyadenosine; G, guanosine 3 AbstractThe formation of 5-methyl-2′-deoxycytidine (5-MedC) following methylation of the C-5 position of cytosine in genomic DNA provides an epigenetic mechanism for the regulation of gene expression and cellular differentiation. We describe the development of a method using HPLC-ultraviolet (UV) detection for the accurate determination of 5-MedC in DNA. Genomic DNA was obtained from HeLa cells and rat liver tissue using an optimised anion-exchange column DNA extraction procedure incorporating a ribonuclease incubation step to remove any potential interference from RNA. Following extraction the DNA samples were enzymatically hydrolysed to 2′-deoxynucleosides using a combination of an endo-exonuclease plus 5′-exonuclease together with a 3′-nucleotidase. The hydrolysed DNA samples (10 µg on column)were analysed using narrow-bore reverse phase HPLC-UV detection. The level of 5-MedC in the DNA samples was expressed as a percentage of the level of 2′-deoxycytidine (dC) determined from calibration lines constructed using authentic standards for 5-MedC and dC. The percentage 5-MedC level determined for commercially available calf thymus DNA was 6.26%, for HeLa cell DNA was 3.02% and for rat liver DNA was 3.55%.4

show abstract

Determination of 5-methylcytosine by acid hydrolysis of DNA with hydrofluoric acid

Cited by 27 publications

References 15 publications

DNA Methylation in Peripheral Blood: A Potential Biomarker for Cancer Molecular Epidemiology

DNA Methylation in Peripheral Blood: A Potential Biomarker for Cancer Molecular Epidemiology

Global DNA methylation measured by liquid chromatography-tandem mass spectrometry: analytical technique, reference values and determinants in healthy subjects

Determination of 5-methyl-2′-deoxycytidine in genomic DNA using high performance liquid chromatography-ultraviolet detection

Contact Info

Product

Resources

About