Abstract
Pulse Field Gel Electrophoresis (PFGE), unlike conventional electrophoresis, can resolve DNA fragments greater than 30 kb and is a highly discriminatory molecular typing method. Here we describe a PFGE protocol for bifidobacteria characterized by a short lysis time determined by the addition of lysis reagents to the initial cell suspension, a reduced incubation period of the plugs in proteinase K, and an improved washing plug step with preheating of the buffer in a shaking incubator.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lee JH, O’Sullivan DJ (2010) Genomic insights into bifidobacteria. Microbiol Mol Biol Rev 74:378–416
Tissier H (1900) Recherches sur la flore intestinale des nourrissons (état normal et pathologique). M.D. thesis. University of Paris, Paris, France
Martín R, Jiménez E, Heilig HG et al (2009) Isolation of bifidobacteria from breast milk and assessment of the bifidobacterial population by PCR-denaturing gradient gel electrophoresis and quantitative real-time PCR. Appl Environ Microbiol 75:965–969
Solís G, de los Reyes-Gavilan CG, Fernández N et al (2010) Establishment and development of lactic acid bacteria and bifidobacteria microbiota in breast-milk and the infant gut. Anaerobe 16:307–310
Martín V, Maldonado A, Moles L et al (2012) Sharing of bacterial strains between breast milk and infant feces. J Hum Lact 28:36–44
Ventura M, O’Flaherty S, Claesson MJ et al (2009) Genome-scale analyses of health-promoting bacteria: probiogenomics. Nat Rev Microbiol 7:61–71
O’Sullivan DJ, Kullen MJ (1998) Tracking of probiotic bifidobacteria in the intestine. Int Dairy J 8:513–525
FAO/WHO (2002) Joint FAO/WHO Working Group report on drafting guidelines for the evaluation of probiotics in food, London, Ontario. World Health Organization, Geneva, Switzerland.
O’Riordan K, Fitzgerald GF (1997) Determination of genetic diversity within the genus Bifidobacterium and estimation of chromosomal size. FEMS Microbiol Lett 156:259–264
Duranti S, Turroni F, Milani C et al (2013) Exploration of the genomic diversity and core genome of the Bifidobacterium adolescentis phylogenetic group by means of a polyphasic approach. Appl Environ Microbiol 79:336–346
McCartney AL, Wenzhi W, Tannock GW (1996) Molecular analysis of the composition of the bifidobacterial and lactobacillus microflora of humans. Appl Environ Microbiol 62:4608–4613
Mättö J, Malinen E, Suihko ML et al (2004) Genetic heterogeneity and functional properties of intestinal bifidobacteria. J Appl Microbiol 97:459–470
Simpson PJ, Stanton C, Fitzgerald GF et al (2003) Genomic diversity and relatedness of bifidobacteria isolated from a porcine cecum. J Bacteriol 185:2571–2581
Grand M, Küffer M, Baumgartner A (2003) Quantitative analysis and molecular identification of bifidobacteria strains in probiotic milk products. Eur Food Res Tech 217:90–92
Gueimonde M, Delgado S, Mayo B et al (2004) Viability and diversity of probiotic Lactobacillus and Bifidobacterium populations included in commercial fermented milks. Food Res Int 37:839–850
Masco L, Huys G, De Brandt E et al (2005) Culture-dependent and culture-independent qualitative analysis of probiotic products claimed to contain bifidobacteria. Int J Food Microbiol 102:221–230
Yeung PSM, Kitts CL, Cano R et al (2004) Application of genotypic and phenotypic analyses to commercial probiotic strain identity and relatedness. J Appl Microbiol 97:1095–1104
Roy D, Ward P, Champagne G (1996) Differentiation of bifidobacteria by use of pulsed-field gel electrophoresis and polymerase chain reaction. Int J Food Microbiol 29:11–29
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Jiménez, E., Gómez, M., Moles, L. (2015). Pulsed Field Gel Electrophoresis for Bifidobacterium . In: Jordan, K., Dalmasso, M. (eds) Pulse Field Gel Electrophoresis. Methods in Molecular Biology, vol 1301. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2599-5_20
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2599-5_20
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2598-8
Online ISBN: 978-1-4939-2599-5
eBook Packages: Springer Protocols