Publication detail

Magnetic hydrophilic microspheres P (HEMA-co-GMA) for DNA isolation from mouse faeces

TRACHTOVÁ, S. OBERMAJER, T. SPANOVA, A. MATIJASIC, B.B. ROGELJ,I. HORAK,D. RITTICH, B.

Original Title

Magnetic hydrophilic microspheres P (HEMA-co-GMA) for DNA isolation from mouse faeces

Type

abstract

Language

English

Original Abstract

Probiotic lactic acid bacteria (LAC) play an important role in health maintenance of the human gastrointestinal tract. A mouse model is widely used for the study of LAC influence on the organism's health. Polymerase chain reaction (PCR) has become a powerful diagnostic tool for the analysis of microorganisms in different types of samples. Faecal samples are a difficult specimen for the application of PCR due to the presence of PCR inhibitors. For this reason an optimal DNA extraction method is crucial for the identification of microorganisms in faecal samples. The problem can be solved by reversible adsorption of whole DNA on magnetic particle surface in the presence of PEG 6000 and sodium chloride. Magnetic non-porous hydrophilic poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) - P(HEMA-co-GMA) microspheres containing carboxyl groups were used for this purpose (Rittich et al. 2009). In this work the proposed method was used for DNA isolation from mouse faeces and identification of Lactobacillus gasseri K7 strain by PCR (Majhenic et al. 2003). The DNA absorption on microsphere surface and the release of adsorbed DNA were optimised. The quality of eluted DNA and the presence of target DNA were examined by PCR and q-PCR using genus-specific Lactobacillus and a specific Lactobacillus gasseri K7 primer set (Dubernet et al. 2002, Majhenic et al. 2003). The next four DNA extraction procedures were used for comparison. It was shown that DNA extracted using P(HEMA-co-EDMA) microspheres is suitable for PCR detection. The amount of isolated DNA was smaller in comparison with the tested procedures but target DNA could be detected. By means of q-PCR no inhibition was detected using undiluted DNA extracts as DNA matrix.

Keywords

Magnetic particles, Lactobacillus gasseri K7

Authors

TRACHTOVÁ, S.; OBERMAJER, T.; SPANOVA, A.; MATIJASIC, B.B.; ROGELJ,I.; HORAK,D.; RITTICH, B.

RIV year

2009

Released

1. 10. 2009

Location

Praha

ISBN

978-80-7080-726-2

Book

Book of Abstract 4th International Symposium on RECENT ADVANCES IN FOOD ANALYSIS

Pages from

259

Pages to

259

Pages count

1

BibTex

@misc{BUT61083,
  author="TRACHTOVÁ, S. and OBERMAJER, T. and SPANOVA, A. and MATIJASIC, B.B. and ROGELJ,I. and HORAK,D. and RITTICH, B.",
  title="Magnetic hydrophilic microspheres P (HEMA-co-GMA) for DNA isolation from mouse faeces",
  booktitle="Book of Abstract 4th International Symposium on RECENT ADVANCES IN FOOD ANALYSIS",
  year="2009",
  pages="259--259",
  address="Praha",
  isbn="978-80-7080-726-2",
  note="abstract"
}