Potential application of superparamagnetic nanoparticles for extraction of bacterial genomic DNA from contaminated food and environmental samples.
BACKGROUND
Isolation of high-molecular-weight DNA is essential for many molecular biology applications. Owing to the presence of polymerase chain reaction (PCR) inhibitors, there is a scarcity of suitable protocols for PCR-ready DNA extraction from food and natural environments. The conventional chemical methods of DNA extraction are time consuming and laborious and the yield is very low. Thus the aim of this research was to develop a simple, rapid, cost-effective method of genomic DNA extraction from food (milk and fruit juice) and environmental (pond water) samples and to detect bacterial contaminants present in those samples.


RESULTS
This approach is efficient for both Gram-positive and Gram-negative bacteria from all the studied samples. Herein super paramagnetic bare iron oxide nanoparticles were implemented for bacterial genomic DNA isolation. The method was also compared to the conventional phenol-chloroform method in the context of quality, quantity and timing process. This method took only half an hour or less to obtain high-molecular-weight purified DNA from minimum bacterial contamination. Additionally, the method was directly compatible to PCR amplification.


CONCLUSIONS
The problem of availability of suitable generalized methods for DNA isolation from various samples including food and environmental has been solved by a nanobiotechnological approach that may prove to be extremely useful in biotechnological applications.
Department of Microbiology, University of Kalyani, West Bengal 741235, India.
Semanti
S
Basu
Saptarshi
S
Chatterjee
Arghya
A
Bandyopadhyay
Keka
K
Sarkar
D000818:Animals; D001419:Bacteria / Q000235:genetics*; D001628:Beverages / Q000382:microbiology*; D002725:Chloroform; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004777:Environment; D005290:Ferric Compounds; D005516:Food Microbiology / Q000379:methods*; D005618:Fresh Water / Q000382:microbiology; D005638:Fruit / Q000382:microbiology; D016680:Genome, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000235:genetics; D059346:Magnets; D008892:Milk / Q000382:microbiology; D008970:Molecular Weight; D053758:Nanoparticles; D010636:Phenols; D016133:Polymerase Chain Reaction; D014871:Water Microbiology
D004269:DNA, Bacterial; D005290:Ferric Compounds; D010636:Phenols; C000499:ferric oxide; D002725:Chloroform