Simple salting-out method for DNA extraction from formalin-fixed, paraffin-embedded tissues

被引:119
作者
Rivero, Elena R. C.
Neves, Adriana C.
Silva-Valenzuela, Maria G.
Sousa, Suzana O. M.
Nunes, Fabio D.
机构
[1] Univ Fed Santa Catarina, Hlth Sci Ctr, Dept Pathol, Florianopolis, SC, Brazil
[2] Butantan Inst, Dept Genet, Sao Paulo, Brazil
[3] Univ Sao Paulo, Sch Dent, Dept Oral Pathol, Sao Paulo, Brazil
关键词
paraffin-embedded tissues; ammonium acetate; salt; DNA extraction; polymerase chain reaction;
D O I
10.1016/j.prp.2006.02.007
中图分类号
R36 [病理学];
学科分类号
100104 [病理学与病理生理学];
摘要
The aim of this study was to standardize a method of DNA extraction from formalin-fixed and paraffin-embedded tissues (PETs) using a salt solution to precipitate protein and isopropanol to precipitate DNA. The samples were submitted to a DNA extraction method in which two different concentrations of ammonium acetate (2 and 4 M) were compared with a phenol-chloroform extraction method and with a commercial DNA isolation kit. DNA was qualified and quantified by spectrophotometer analysis, electrophoresis, and amplification by PCR. The 167 and 268 bp fragments of APC and beta-globin genes, respectively, were amplified equally from DNA extracted by all tested methods and in all cases. However, the 536 bp fragment of beta-globin gene was not amplified in all cases. According to our results, the extraction method using ammonium acetate proved to be simple and suitable for obtaining DNA of good quality, which can be easily amplified by PCR. (c) 2006 Elsevier GmbH. All rights reserved.
引用
收藏
页码:523 / 529
页数:7
相关论文
共 27 条
[1]
Salvia columbariae contains tanshinones [J].
Adams, JD ;
Wall, M ;
Garcia, C .
EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE, 2005, 2 (01) :107-110
[2]
AKAO I, 1991, LARYNGOSCOPE, V101, P279
[3]
BANERJEE SK, 1995, BIOTECHNIQUES, V18, P768
[4]
Comparison of methods for DNA extraction from paraffin-embedded tissues and buccal cells [J].
Cao, W ;
Hashibe, M ;
Rao, JY ;
Morgenstern, H ;
Zhang, ZF .
CANCER DETECTION AND PREVENTION, 2003, 27 (05) :397-404
[5]
SIMPLIFIED PROCEDURES FOR APPLYING THE POLYMERASE CHAIN-REACTION TO ROUTINELY FIXED PARAFFIN WAX SECTIONS [J].
COATES, PJ ;
DARDENNE, AJ ;
KHAN, G ;
KANGRO, HO ;
SLAVIN, G .
JOURNAL OF CLINICAL PATHOLOGY, 1991, 44 (02) :115-118
[6]
DABREK J, 2002, BIOMED PAP, V146, P37
[7]
DNA extraction from formalin-fixed, paraffin-embedded tissues: Protein digestion as a limiting step for retrieval of high-quality DNA [J].
Diaz-Cano, SJ ;
Brady, SP .
DIAGNOSTIC MOLECULAR PATHOLOGY, 1997, 6 (06) :342-346
[8]
EFFECTS OF FIXATIVE AND FIXATION TIME ON THE EXTRACTION AND POLYMERASE CHAIN-REACTION AMPLIFICATION OF RNA FROM PARAFFIN-EMBEDDED TISSUE - COMPARISON OF 2 HOUSEKEEPING GENE MESSENGER-RNA CONTROLS [J].
FOSS, RD ;
GUHATHAKURTA, N ;
CONRAN, RM ;
GUTMAN, P .
DIAGNOSTIC MOLECULAR PATHOLOGY, 1994, 3 (03) :148-155
[9]
PURIFICATION OF DNA FROM FORMALDEHYDE FIXED AND PARAFFIN EMBEDDED HUMAN-TISSUE [J].
GOELZ, SE ;
HAMILTON, SR ;
VOGELSTEIN, B .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1985, 130 (01) :118-126
[10]
PCR AMPLIFICATION FROM PARAFFIN-EMBEDDED TISSUES - EFFECTS OF FIXATIVE AND FIXATION TIME [J].
GREER, CE ;
PETERSON, SL ;
KIVIAT, NB ;
MANOS, MM .
AMERICAN JOURNAL OF CLINICAL PATHOLOGY, 1991, 95 (02) :117-124