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doi: 10.1016/S0001-6519(04)78497-6
Extracción de adn con resina chelex en el análisis de la amplificación oncogénica en carcinomas de cabeza y cuello
DNA extraction using chelex resin for the oncogenic amplification analysis in nead and neck tumours
L.A. García González,a, , J.P. Rodrigo Tapia**, P. Sánchez Lazo***, S. Ramos***, C. Suárez Nieto**
* Servicio de orl. hospital carmen y severo ochoa. asturias
** Servicio de orl. hospital central de asturias. instituto de oncología. universidad de oviedo
*** Departamento de biología molecular. instituto de oncología. universidad de oviedo
Recibido 10 diciembre 2003, Aceptado 29 enero 2004
Resumen

La extracción de ADN de tejido tumoral puede ser el proceso más laborioso y complejo en la amplificación de ADN mediante PCR cuando se utiliza el procedimiento con fenol-cloroformo. Comparamos este método de extracción extenso, lento y caro con otras dos técnicas basadas en el uso de resina Chelex-100. Esta resina quelante ha sido utilizada para la extracción de ADN de diferentes tejidos para su uso con la PCR. Estos procedimientos son simples, rápidos y no requieren múltiples pasos. En este trabajo comparamos la extracción de ADN procedente de 30 carcinomas epidermoides de cabeza y cuello utilizando la precipitación con solventes orgánicos, resina Chelex-100 con y sin procesamiento previo con proteinasa K. Los resultados evidencian que el procedimiento con proteinasa K y resina Chelex-100 es un método tan eficaz como la precipitación con fenol-cloroformo

Abstract

DNA extraction from tissues can be the most laborious and complex step in amplifying DNA by PCR when phenol-choroform procedure is used. We compare this lengthy, slow and expensive extraction method with other two based in the use of Chelex-100 resin. This chelating resin has been applied for extracting DNA from different tissues to use with the PCR. These procedures are simple, rapid and do not require multiple steps. In this study we compared DNA extraction from 30 head and neck squamous cell carcinomas (HNSCC) using organic solvent precipitation, Chelex 100 resin with and without proteinase K pretreatment. The results show that proteinase K-Chelex 100 procedure is as efficient as the phenol-chloroform one

Palabras clave
Tumores de cabeza y cuello, Factores de crecimiento fibroblástico, Extracción de ADN, Amplificación de oncogenes, Reacción en cadena de polimerasa, Resina Chelex-100
Key words
Head cancer, Neck cancer, Fibroblastic growth factors, DNA extraction, Oncogene amplification, Polymerase chain reaction, Chelex 100-resin
El Texto completo solo esta disponible en PDF
Referencias
1.
N. Lench,P. Stanier,R. Williamson
Simple non-invasive method to obtain DNA for gene analysis
Lancet, 8599 (1988), pp. 1356-1358
2.
D.N. Miller,J.E. Bryant,E.L. Madse,W.C. Ghiorse
Evaluation and optimization of DNA extraction and purification procedures for soil and sediment samples
Appl Environ Microbiol, 65 (1999), pp. 4715-4724
3.
M.J. Heller,R.A. Robinson,L.J. Burgart,C.J. TenEyck,W.W. Wilke
DNA extraction by sonication: a comparison of fresh, frozen, and paraffin-embedded tissues extracted for use in polymerase chain reaction assays
Mod Pathol, 5 (1992), pp. 203-206
4.
P.S. Walsh,D.A. Metzger,R. Higuchi
Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material
Biotechniques, 10 (1991), pp. 506-513
5.
C. Zandotti,X. de Lamballerie,C. Guignole-Vignoli,C. Bollet,P. de Micco
A Rapid DNA extraction method from culture -. and clinical samples. Suitable for the detection of human cytomegalovirus by the polymerase chain reaction
Acta Virol, 37 (1993), pp. 106-108
6.
H. Ellegren
Genomic DNA from museum bird feathers
Ancient DNA, pp. pp 211-pp 217
7.
R. Sepp,I. Szabó,H. Uda,H. Sakamoto
Rapid techniques for DNA extraction from routinely processed archival tissue for use in PCR
J Clin Pathol, 47 (1994), pp. 318-323
8.
C. Vignoli,X. de Lamallerie,C. Zandotti,C. Tamalet,P. de Micco
Advantage of a rapid extraction method of HIV1 DNA suitable for polymerase chain reaction
Res Virol, 146 (1995), pp. 159-162
9.
A. Estoup,C.R. Largiadèr,E. Perrot,D. Chourront
Rapid one-tube DNA extraction for reliable PCR detection of fish polymorphic markers and transgenes
Molecular Marine Biology and Biotechnology, 5 (1996), pp. 295-298
10.
P. Gill,C.P. Kimpton,K. Sullivan
A rapid polymerase chain reaction method for identifying fixed specimens
Electrophoresis, 13 (1992), pp. 173-175
11.
J.P. Rodrigo,L.A. García,P.S. Lazo,S. Ramos,C. Suárez
PCR analysis of INT-2 oncogene amplification in squamous cell carcinomas of the head and neck
Otolaryngol. Head Neck Surg, 115 (1996), pp. 82
12.
J. Sambrook,E.F. Fritsch,T. Maniatis
Cold Spring Harbor Laboratory Press, (1989)
13.
J.P. Rodrigo Tapia,L.A. García,P. Sánchez Lazo,S. Ramos,C. Suárez Nieto
EMS1 gene amplification correlates with poor prognosis in squamous cell carcinomas of the head and neck
Clinical Cancer Research, 6 (2000), pp. 3177-3182
14.
G. Giraffa,L. Rossetti,E. Neviani
An evaluation of chelex-based DNA purification protocols for the typing of lactic acid bacteria
J Microbiol Methods, 42 (2000), pp. 175-184
15.
J. Singer-Sam,R.L. Tanguay,A.D. Riggs
Use of Chelex to improve the PCR signal from a small number of cells
Amplifications, 3 (1989), pp. 11
16.
D.K. Wright,M.M. Manos
Sample preparation from paraffin.embedded tissues
PCR protocols: a guide to methods and applications, pp. 153-156
17.
J. Attal,M. Cajero-Juarez,L.M. Houdebine
A simple method of DNA extraction from whole tissues and blood using glass powder for detection of transgenic animals by PCR
Transgenis Res, 4 (1995), pp. 149-150
18.
A. Vince,M. Poljak,K. Seme
DNA extraction from archival Giemsa-stained bone-marrow slides: comparison of six rapid methods
Br J Haematol, 101 (1998), pp. 349-351
19.
J.M. Jung,C.T. Comey,D.B. Baer,B. Budowle
Extraction strategy for obtaining DNA from bloodsatins for PCR amplification and typing of the HLA-DQ alpha gene
Int J Legal Med, 104 (1991), pp. 145-148
20.
N. Vandenberg,R.A. van Oorschot,R.J. Mitchell
An evaluation of selected DNA extraction strategies for short tandem repeat typing
Electrophoresis, 18 (1997), pp. 1624-1626
21.
X. de Llamballerie,C. Zandotti,C. Vignoli,C. Bollet,P. de Micco
A one-step microbial DNA extraction method using “Chelex 100” suitable for gene amplification
Res Microbiol, 143 (1992), pp. 785-790
22.
C. Muñoz,M. Jane,A. González-Cuevas,T. Juncosa,A. Gene,V. Varea
Evaluation of a simple rapid polymerase chain reaction (PCR) technique for the diagnosis of Helicobacter pylori infection in childhood
Enferm Infecc Microbiol Clin, 17 (1999), pp. 119-125
23.
P. Mohlenhoff,L. Muller,A.A. Gorbushina,K. Petersen
Molecular approach to the characterisation of fungal communities: methods for DNA extraction, PCR amplification and DGGE analysis of painted art objects
FEMS Microbiol Letts, 195 (2001), pp. 169-173
24.
M. Panaccio,M. Georgesz,C. Hollywell,A. Lew
Direct PCR from solid tissues without DNA extraction
Nucleic Acids Res, 21 (1993), pp. 4656
25.
Y.S. Liu,R.J. Thomas,W.A. Phillips
Single-step direct PCR amplification from solid tissues
Nucleic Acids Res, 23 (1995), pp. 1640
26.
N.J. Coombs,A.C. Gough,J.N. Primrose
Optimisation of DNA and RNA extraction from archival formalin-fixed tissue
Nucleic Acids Res, 27 (1999), pp. e12
27.
A. Ohhashi,T. Aoki,S. Matsugo,C. Simasaki
PCR-based typing of human buccal cell´s DNA extracted from whole saliva and saliva stains
Nippon Hiogaku Zasshi, 47 (1993), pp. 108-118
Correspondencia: Servicio de ORL. Hospital Carmen y Severo Ochoa. Sienra, 11. Cangas del Narcea. 33800 Asturias
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