Submitted: 16 Aug 2015
Revised: 06 Sep 2015
Accepted: 22 Sep 2015
First published online: 06 Oct 2016
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Int J Enteric Pathog. 2016;4(1): e32472.
doi: 10.17795/ijep32472
  Abstract View: 910
  PDF Download: 572

Research Article

An Efficient DNA Extraction Method for Lactobacillus casei, a Difficult-to-Lyse Bacterium

Mojtaba Alimolaei 1,2, Mehdi Golchin 1 *

1 Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, IR Iran
2 Department of Anaerobic Bacterial Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Kerman Branch, Kerman, IR Iran
*Corresponding author: Mehdi Golchin, Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, IR Iran. Tel/Fax: +98-343257447, Email: golchin@uk.ac.ir

Abstract

Background: There are several protocols to extract DNA from Lactobacillus spp. In the case of L. casei it is harder because of its especial and thick cell wall.

Objectives: In this study, nine DNA extraction protocols (by lysozyme treatment) were evaluated and compared in two categories (traditional and kit-based protocols) and an improved method was presented.

Materials and Methods: DNA quantity and quality was determined by spectrophotometry, agarose gel electrophoresis and polymerase chain reaction (PCR).

Results: The results revealed that the yield of extracted DNA differed by each protocol (5.8 - 17.1 μg/100 μL), but provided appropriate DNA for PCR amplification. The modified protocol offered the best total DNA extraction method when both quality (DNA purity; 1.54 μg) and quantity (DNA yield; 17.1 μg) were considered.

Conclusions: We suggest this protocol for effective and inexpensive DNA isolation from L. casei for downstream biological processes such as PCR.

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