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The evaluation of the microbial diversity in anthropogenic subtrates using a method analyzing the polymorphism of the terminal restriction fragments of DNA
Gryndler, Milan; Bukovská, Petra; Sýkorová, Zuzana; Havránková, Marie
2010 - Czech
In this methodology we describe a fast, easy and non-expensive method for evaluation of microbial diversity in various types of substrates: T-RFLP. We explain principles and describe in detail the workflow of DNA extraction, amplification of a particular piece of DNA in PCR, choice of restriction enzymes and the restriction digest, separation of fragments using the capillary sequencer as well as discuss the evaluation and interpretation of the results. V této metodice popisujeme rychlou, snadnou a nepříliš nákladnou metodu vyhodnocení mikrobiální diverzity v různých typech substrátů: T-RFLP. Popisujeme principy a detailně vysvětlujeme postup práce při extrakci DNA, amplifikaci určitého úseku DNA pomocí PCR, při výběru restrikčního enzymu a restrikčním štěpení, dělení fragmentů v kapilárním sekvenátoru a rovněž diskutujeme vyhodnocení a interpretaci výsledků.
Keywords:
t-RFLP; microorganism; diversity
Available at various institutes of the ASCR
The evaluation of the microbial diversity in anthropogenic subtrates using a method analyzing the polymorphism of the terminal restriction fragments of DNA
In this methodology we describe a fast, easy and non-expensive method for evaluation of microbial diversity in various types of substrates: T-RFLP. We explain principles and describe in detail the ...
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