Abstract
A simple and inexpensive MEKC method, which is able to assess base damage within DNA samples, is illustrated. After heat-acid hydrolysis of the DNA samples, both the percentage of each canonical DNA base and the relative amount of uncanonical DNA bases can be measured. This method is useful for an evaluation of the integrity of PCR templates used in several fields of investigation.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lindahl T (1993) Instability and decay of the primary structure of DNA. Nature 362:709–715
De Bont R, Lorebeke N (2004) Endogenous DNA damage in humans: a review of quantitative data. Mutagenesis 19:169–185
Dizdaroglu M (1985) Application of capillary gas-chromatography mass-spectrometry to chemical characterization of radiation-induced base damage of DNA: implications for assessing DNA-repair processes. Anal Biochem 144:593–603
Lassen C, Hummel S, Herrmann B (1984) Comparison of DNA extraction and amplification from ancient human bone and mummified soft tissue. Int J Leg Med 107:152–155
Pääbo S (1989) Ancient DNA: extraction, characterization, molecular cloning, and enzymatic amplification. Proc Natl Acad Sci USA 86:1939–1943
Höss M, Jaruga P, Zastawny TH, Dizdaroglu M, Pääbo S (1996) DNA damage and DNA sequence retrieval from ancient tissues. Nucleic Acids Res 24:1304–1307
Fattorini P, Ciofuli R, Cossutta F, Giulianini P, Edomi P, Furlanut M, Previderé C (1999) Fidelity of polymerase chain reaction-direct sequencing analysis of damaged forensic Âsamples. Electrophoresis 20:3349–3357
Previderè C, Micheletti P, Perossa R, Grignani P, Fattorini P (2002) Molecular Âcharacterisation of the nucleic acids recovered from aged forensic samples. Int J Legal Med 116:334–339
Zimmermann J, Hajibabaei M, Blackburn DC, Hanken J, Cantin E, Posfai J, Evans TC Jr (2008) DNA damage in preserved specimens and tissue samples: a molecular assessment. Front Zool 23:5–18
Pizzichini M, Arezzini L, Billarelli C, Carlucci F, Terzuoli L (1998) Determination and separation of allantoin, uric acid, hypoxanthine, and xanthine by capillary zone electrophoresis. Adv Exp Med Biol 431:797–800
Alfazema LN, Howells S, Perrett D (1998) Determination of allantoin in biofluids using micellar electrokinetic capillary chromatography. J Chromatogr A 817:345–352
Adam T, Friedecký D, Fairbanks LD, SevcÃk J, Barták P (1999) Capillary electrophoresis for detection of inherited disorders of purine and pyrimidine metabolism. Clin Chem 45:2086–2093
Friedecký D, Adam T, Barták P (2002) Capillary electrophoresis for detection of inherited disorders of purine and pyrimidine metabolism: a selective approach. Electrophoresis 23:565–571
Esaka Y, Inagaki S, Goto M (2003) Separation procedures capable of revealing DNA adducts. J Chromatogr B Analyt Technol Biomed Life Sci Chromatogr 797:321–329
Ma Y, Liu G, Du M, Stayton I (2004) Recent developments in the determination of urinary cancer biomarkers by capillary electrophoresis. Electrophoresis 25:1473–1484
Zhang L, Liu Y, Chen G (2004) Simultaneous determination of allantoin, choline and l-arginine in Rhizoma Dioscoreae by capillary electrophoresis. J Chromatogr A 1043:317–321
Cheng ML, Liu TZ, Lu FJ, Chiu DTY (1999) Simultaneous detection of vitamin C and uric acid by capillary electrophoresis in plasma of diabetes and in aqueous humor in acute Âanterior uveitis. Clin Biochem 32:473–476
Terzuoli L, Porcelli B, Setacci C, Giubbolini M, Cinci G, Carlucci F, Pagani R, Marinello E (1999) Comparative determination of purine compounds in carotid plaque by capillary zone electrophoresis and high-performance liquid chromatography. J Chromatogr B 728:185–192
Carlucci F, Tabucchi A, Biagioli B, Sani G, Lisi G, Maccherini M, Rosi F, Marinello E (2000) Capillary electrophoresis in the evaluation of ischemic injury: simultaneous determination of purine compounds and glutathione. Electrophoresis 21:1552–1557
Zinellu A, Carru C, Sotgia S, Deiana L (2004) Optimization of ascorbic and uric acid separation in human plasma by free zone capillary electrophoresis ultraviolet detection. Anal Biochem 330:298–305
Kattygnarath D, Mounier N, Madelaine-Chambrin I, Gourmel B, Le Bricon T, Gisselbrecht C, Faure P, Houze P (2006) Quantification of urinary allantoin by capillary zone electrophoresis during recombinant urate oxydase (rasburicase) therapy. Clin Biochem 39:86–90
Silva M (2011) Micellar electrokinetic chromatography: a practical overview of current methodological and instrumental advances. Electrophoresis 32:149–165
Fattorini P, Marrubini G, Ricci U, Gerin F, Grignani P, Sorçaburu Cigliero S, Xamin A, Edalucci E, La Marca G, Previderé C (2009) Estimating the integrity of aged DNA samples by CE. Electrophoresis 30:3986–3995
Fattorini P, Marrubini G, Sorçaburu Cigliero S, Pitacco P, Grignani P, Previderè C (2011) CE analysis and molecular characterization of depurinated DNA samples. Electrophoresis 32:3042–3052
Araujo P (2009) Key aspects of analytical method validation and linearity evaluation. J Chromatogr B 877:2224–2234
Acknowledgments
The authors wish to thank Dr. Sergio G. Tisminetzky and C. Guarnaccia (ICGEB, Trieste, Italy) for their invaluable support in this work.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Fattorini, P., Marrubini, G., Grignani, P., Sorçaburu-Cigliero, S., Previderé, C. (2013). Assessment of DNA Damage by Micellar Electrokinetic Chromatography. In: Volpi, N., Maccari, F. (eds) Capillary Electrophoresis of Biomolecules. Methods in Molecular Biology, vol 984. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-296-4_25
Download citation
DOI: https://doi.org/10.1007/978-1-62703-296-4_25
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-295-7
Online ISBN: 978-1-62703-296-4
eBook Packages: Springer Protocols