Determination of 5-methylcytosine by acid hydrolysis of DNA with hydrofluoric acid
Quantitation of 5-methylcytosine in DNA after acid hydrolysis has been inaccurate because deamination of cytosine and 5-methylcytosine occurs during the hydrolysis procedure. There is little information in the literature regarding the use of hydrofluoric acid (HF) for DNA hydrolysis and we have ther...
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| Veröffentlicht in: | Analytical biochemistry 1987-12, Vol.167 (2), p.347-351 |
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| container_title | Analytical biochemistry |
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| creator | Catania, J. Keenan, B.C. Margison, G.P. Fairweather, D.S. |
| description | Quantitation of 5-methylcytosine in DNA after acid hydrolysis has been inaccurate because deamination of cytosine and 5-methylcytosine occurs during the hydrolysis procedure. There is little information in the literature regarding the use of hydrofluoric acid (HF) for DNA hydrolysis and we have therefore undertaken a systematic study of this process. The deoxyribonucleotides of cytosine and 5-methylcytosine were shown not to undergo detectable levels of deamination during prolonged periods (up to 24 h) at 80°C in 48% HF. Kinetic studies show that the release of purine and pyrimidine bases was complete by 4 h under these conditions. Analysis of the 5-methylcytosine content of DNA from various tissues gave levels that were very close to the values reported in the literature. This method is ideally suited for the determination of the overall cytosine methylation levels in DNA. |
| doi_str_mv | 10.1016/0003-2697(87)90175-8 |
| format | Article |
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There is little information in the literature regarding the use of hydrofluoric acid (HF) for DNA hydrolysis and we have therefore undertaken a systematic study of this process. The deoxyribonucleotides of cytosine and 5-methylcytosine were shown not to undergo detectable levels of deamination during prolonged periods (up to 24 h) at 80°C in 48% HF. Kinetic studies show that the release of purine and pyrimidine bases was complete by 4 h under these conditions. Analysis of the 5-methylcytosine content of DNA from various tissues gave levels that were very close to the values reported in the literature. This method is ideally suited for the determination of the overall cytosine methylation levels in DNA.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/0003-2697(87)90175-8</identifier><identifier>PMID: 3442330</identifier><identifier>CODEN: ANBCA2</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>5-Methylcytosine ; acid hydrolysis ; Analytical, structural and metabolic biochemistry ; Animals ; Base Composition ; Biological and medical sciences ; Chromatography, High Pressure Liquid ; Cytosine - analogs & derivatives ; Cytosine - analysis ; Deamination ; DNA - analysis ; Fundamental and applied biological sciences. Psychology ; HPLC ; Hydrofluoric Acid ; Hydrolysis ; kinetics ; Methylation ; Nucleic acids ; Nucleic bases, nucleotides</subject><ispartof>Analytical biochemistry, 1987-12, Vol.167 (2), p.347-351</ispartof><rights>1987</rights><rights>1989 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-38571bc68dc4253417772a40ddc17def6869863764b55968fa36e95e28bc51963</citedby><cites>FETCH-LOGICAL-c417t-38571bc68dc4253417772a40ddc17def6869863764b55968fa36e95e28bc51963</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0003-2697(87)90175-8$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3538,27906,27907,45977</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7014519$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3442330$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Catania, J.</creatorcontrib><creatorcontrib>Keenan, B.C.</creatorcontrib><creatorcontrib>Margison, G.P.</creatorcontrib><creatorcontrib>Fairweather, D.S.</creatorcontrib><title>Determination of 5-methylcytosine by acid hydrolysis of DNA with hydrofluoric acid</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>Quantitation of 5-methylcytosine in DNA after acid hydrolysis has been inaccurate because deamination of cytosine and 5-methylcytosine occurs during the hydrolysis procedure. There is little information in the literature regarding the use of hydrofluoric acid (HF) for DNA hydrolysis and we have therefore undertaken a systematic study of this process. The deoxyribonucleotides of cytosine and 5-methylcytosine were shown not to undergo detectable levels of deamination during prolonged periods (up to 24 h) at 80°C in 48% HF. Kinetic studies show that the release of purine and pyrimidine bases was complete by 4 h under these conditions. Analysis of the 5-methylcytosine content of DNA from various tissues gave levels that were very close to the values reported in the literature. This method is ideally suited for the determination of the overall cytosine methylation levels in DNA.</description><subject>5-Methylcytosine</subject><subject>acid hydrolysis</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Base Composition</subject><subject>Biological and medical sciences</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Cytosine - analogs & derivatives</subject><subject>Cytosine - analysis</subject><subject>Deamination</subject><subject>DNA - analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>HPLC</subject><subject>Hydrofluoric Acid</subject><subject>Hydrolysis</subject><subject>kinetics</subject><subject>Methylation</subject><subject>Nucleic acids</subject><subject>Nucleic bases, nucleotides</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtr3DAQgEVpSDdp_0ELPpSSHpyOrPclEJI-AiGF0p6FLI1ZFdtKJW-L_33t7LLH9jQw882Dbwh5TeGSApUfAIDVjTTqQqv3BqgStX5GNhSMrIGBeU42R-QFOSvlJwClXMhTcso4bxiDDfl2ixPmIY5uimmsUleJesBpO_d-nlKJI1btXDkfQ7WdQ079XGJZsduH6-pPnLb7dNfvUo7-CXxJTjrXF3x1iOfkx6eP32--1PdfP9_dXN_XnlM11UwLRVsvdfC8EWzJKdU4DiF4qgJ2UkujJVOSt0IYqTvHJBqBjW69oEayc_JuP_cxp187LJMdYvHY927EtCtWKSOBM_VfkApgWhuzgHwP-pxKydjZxxwHl2dLwa7O7SrUrkKtVvbJudVL25vD_F07YDg2HSQv9beHuive9V12o4_liClYnkLX7Vd7DBdpvyNmW3zE0WOIGf1kQ4r_vuMv00ibnA</recordid><startdate>19871201</startdate><enddate>19871201</enddate><creator>Catania, J.</creator><creator>Keenan, B.C.</creator><creator>Margison, G.P.</creator><creator>Fairweather, D.S.</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>19871201</creationdate><title>Determination of 5-methylcytosine by acid hydrolysis of DNA with hydrofluoric acid</title><author>Catania, J. ; Keenan, B.C. ; Margison, G.P. ; Fairweather, D.S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-38571bc68dc4253417772a40ddc17def6869863764b55968fa36e95e28bc51963</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>5-Methylcytosine</topic><topic>acid hydrolysis</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Base Composition</topic><topic>Biological and medical sciences</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Cytosine - analogs & derivatives</topic><topic>Cytosine - analysis</topic><topic>Deamination</topic><topic>DNA - analysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>HPLC</topic><topic>Hydrofluoric Acid</topic><topic>Hydrolysis</topic><topic>kinetics</topic><topic>Methylation</topic><topic>Nucleic acids</topic><topic>Nucleic bases, nucleotides</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Catania, J.</creatorcontrib><creatorcontrib>Keenan, B.C.</creatorcontrib><creatorcontrib>Margison, G.P.</creatorcontrib><creatorcontrib>Fairweather, D.S.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Catania, J.</au><au>Keenan, B.C.</au><au>Margison, G.P.</au><au>Fairweather, D.S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Determination of 5-methylcytosine by acid hydrolysis of DNA with hydrofluoric acid</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>1987-12-01</date><risdate>1987</risdate><volume>167</volume><issue>2</issue><spage>347</spage><epage>351</epage><pages>347-351</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><coden>ANBCA2</coden><abstract>Quantitation of 5-methylcytosine in DNA after acid hydrolysis has been inaccurate because deamination of cytosine and 5-methylcytosine occurs during the hydrolysis procedure. There is little information in the literature regarding the use of hydrofluoric acid (HF) for DNA hydrolysis and we have therefore undertaken a systematic study of this process. The deoxyribonucleotides of cytosine and 5-methylcytosine were shown not to undergo detectable levels of deamination during prolonged periods (up to 24 h) at 80°C in 48% HF. Kinetic studies show that the release of purine and pyrimidine bases was complete by 4 h under these conditions. Analysis of the 5-methylcytosine content of DNA from various tissues gave levels that were very close to the values reported in the literature. This method is ideally suited for the determination of the overall cytosine methylation levels in DNA.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>3442330</pmid><doi>10.1016/0003-2697(87)90175-8</doi><tpages>5</tpages></addata></record> |
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| ispartof | Analytical biochemistry, 1987-12, Vol.167 (2), p.347-351 |
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| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | 5-Methylcytosine acid hydrolysis Analytical, structural and metabolic biochemistry Animals Base Composition Biological and medical sciences Chromatography, High Pressure Liquid Cytosine - analogs & derivatives Cytosine - analysis Deamination DNA - analysis Fundamental and applied biological sciences. Psychology HPLC Hydrofluoric Acid Hydrolysis kinetics Methylation Nucleic acids Nucleic bases, nucleotides |
| title | Determination of 5-methylcytosine by acid hydrolysis of DNA with hydrofluoric acid |
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