![]() Variations in the composition of the gut microbiota between individuals are increasingly recognized due to the development of culture-independent analytic techniques ( Smith et al., 2011). DGGE and T-RFLP produced the highest diversity scores for DNA extracted using kit Z (H′ = 2.30 and 1.27) and kit QS (H′ = 2.16 and 0.94), which also extracted the highest DNA yields compared to the other kits assessed. ![]() qPCR results showed that all kits were uniformly efficient for extracting DNA from the selected target bacteria. All kits were shown to be reproducible with CV values ≤ 0.46 for DNA extraction. Automated kit QS exhibited the best quality and highest quantity of DNA. The measurements of DNA yield and purity exhibited variations between the five kits tested in this study. Three bacteria, commonly found in human faeces were quantified using real time polymerase chain reaction (qPCR) and total bacterial diversity was studied using denaturing gradient gel electrophoresis (DGGE) as well as terminal restriction fragment length polymorphism (T-RFLP). Yield, purity and integrity of total genomic DNA were compared spectrophotometrically and using gel electrophoresis. Five widely used commercial deoxyribonucleic acid (DNA) extraction kits (QIAsymphony® Virus/Bacteria Midi Kit (kit QS), ZR Fecal DNA MiniPrep™ (kit Z), QIAamp® DNA Stool Mini Kit (kit QA), Ultraclean® Fecal DNA Isolation Kit (kit U) and PowerSoil® DNA Isolation Kit (kit P)) were evaluated, using human faecal samples. Reliable extraction of nucleic acid is a key step in identifying the composition of the faecal microbiota. Differences in the composition of the gut microbiota have been associated with a range of diseases using culture-independent methods.
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