An evaluation of chelex-based DNA purification protocols for the typing of lactic acid bacteria

被引：113

作者：

Giraffa, G ^{[1
]}

Rossetti, L ^{[1
]}

Neviani, E ^{[1
]}

机构：

[1] Ist Sperimentale Lattiero Caseario, I-26900 Lodi, Italy

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 2000年 / 42卷 / 02期

关键词：

bacterial taxonomy; diagnostic microbiology; DNA extraction; polymerase chain reaction (PCR) fingerprinting; lactic acid bacteria;

D O I：

10.1016/S0167-7012(00)00172-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An easy and rapid protocol to extract DNA to be used as template for polymerase chain reaction (PCR) fingerprinting experiments from cultivable lactic acid bacteria (LAB) is proposed. Different procedures for rapid extraction of DNA by chelex (iminodiacetid acid) ionic resin were compared. Factors affecting the quality and reproducibility of PCR fingerprinting profiles were also investigated. Two out of three chelex-based protocols allowed to obtain DNA samples which, after PCR amplification, provided electrophoretic patterns comparable with those obtained by classical lysozyme and phenol-chloroform DNA extraction. A good level of reproducibility and consistency of the InstaGene procedure was verified. The procedure is fast, practical, and the DNA is of quality similar to that obtained by phenol-chloroform extraction. Although applied to a little number of LAB strains, chelex-based protocols are potentially applicable to a vast array of organisms and/or biological materials. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：175 / 184

页数：10

共 25 条

[1] Molecular identification and cluster analysis of homofermentative thermophilic lactobacilli isolated from dairy products [J].

Andrighetto, C ;

De Dea, P ;

Lombardi, A ;

Neviani, E ;

Rossetti, L ;

Giraffa, G .

RESEARCH IN MICROBIOLOGY, 1998, 149 (09) :631-643

[2]

[Anonymous], CHEM METHODS PROKARY

[3] DEVELOPMENT OF RAPD PROTOCOL FOR TYPING OF STRAINS OF LACTIC-ACID BACTERIA AND ENTEROCOCCI [J].

COCCONCELLI, PS ;

PORRO, D ;

GALANDINI, S ;

SENINI, L .

LETTERS IN APPLIED MICROBIOLOGY, 1995, 21 (06) :376-379

[4]

Corroler D, 1998, APPL ENVIRON MICROB, V64, P4729

[5] THE EFFECT OF COLONY AGE ON PCR FINGERPRINTING [J].

COUTINHO, HLC ;

HANDLEY, BA ;

KAY, HE ;

STEVENSON, L ;

BERINGER, JE .

LETTERS IN APPLIED MICROBIOLOGY, 1993, 17 (06) :282-284

[6] Differentiation of Lactobacillus helveticus strains using molecular typing methods [J].

Drake, MA ;

Small, CL ;

Spence, KD ;

Swanson, BG .

FOOD RESEARCH INTERNATIONAL, 1996, 29 (5-6) :451-455

[7] STRAIN TYPING IN THE GENUS LACTOBACILLUS [J].

DYKES, GA ;

VONHOLY, A .

LETTERS IN APPLIED MICROBIOLOGY, 1994, 19 (02) :63-66

[8] HYPERVARIABLE DNA FINGERPRINTING IN ESCHERICHIA-COLI - MINISATELLITE PROBE FROM BACTERIOPHAGE-M13 [J].

HUEY, B ;

HALL, J .

JOURNAL OF BACTERIOLOGY, 1989, 171 (05) :2528-2532

[9] Taxonomy and physiology of probiotic lactic acid bacteria [J].

Klein, G ;

Pack, A ;

Bonaparte, C ;

Reuter, G .

INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 1998, 41 (02) :103-125

[10]

Moschetti G, 1998, J APPL MICROBIOL, V85, P25, DOI 10.1046/j.1365-2672.1998.00461.x

← 1 2 3 →