An evaluation of chelex-based DNA purification protocols for the typing of lactic acid bacteria.
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.
Giraffa
G
G
Istituto Sperimentale Lattiero Caseario, Via A. Lombardo 11, 26900 Lodi, Italy. giraffa@pop.telware.it
Rossetti
L
L
Neviani
E
E
D015373:Bacterial Typing Techniques; D002725:Chloroform / Q000737:chemistry; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000302:isolation & purification*; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics; D013294:Lactococcus lactis / Q000145:classification* / Q000235:genetics; D019800:Phenol / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011137:Polystyrenes; D011145:Polyvinyls; D015203:Reproducibility of Results
D004269:DNA, Bacterial; D011137:Polystyrenes; D011145:Polyvinyls; D019800:Phenol; D002725:Chloroform; C006960:chelex