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The first decade of terminal restriction fragment length polymorphism (T-RFLP) in microbial ecology
Stres, Blaž (Author)

URLURL - Presentation file, Visit http://www.dlib.si/details/URN:NBN:SI:doc-Q6NNUIB2 New window

Abstract
Terminal restriction fragment length polymorphism (T-RFLP) was introduced to environmental microbiology only a decade ago but it soon became a molecular tool of choice, due to its high throughput and phylogenetic resolution. Fierce discussions accompanied the new method leading to sophistication of the data preparation, acquisition, manipulation and standardization of analysis. Consequently, numerous approaches were proposed at various steps and also criticized. As a result, a combination of variable percentage threshold and Bray-Curtis index used in non-metric multidimensional scaling are now being accepted. Their combination offers a balance between noise elimination and information retention yielding a powerful and yet easily interpreted method toexamine community patterns based on T-RFLP data. Its current state of the art and future developments highlight the potential of the method in the field of microbial ecology. However, a more standardized approach and a higher level of control at all stages of T-RFLP fingerprinting are needed.

Language:English
Keywords:microbiology, microbial ecology, molecular biology, molecular genetics, terminal restriction fragment length polymorphism, T-RFLP
Work type:Not categorized (r6)
Tipology:1.02 - Review Article
Organization:BF - Biotechnical Faculty
Year:2006
Publisher:Biotehniška fakulteta
Number of pages:str. 65-73
Numbering:Letn. 88, št. 2
UDC:579
ISSN on article:1581-9175
COBISS.SI-ID:1971592 Link is opened in a new window
Views:416
Downloads:127
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Record is a part of a journal

Title:Acta agriculturae Slovenica
Shortened title:Acta agric. Slov.
Publisher:Biotehniška fakulteta
ISSN:1581-9175
COBISS.SI-ID:213840640 New window

Secondary language

Language:Slovenian
Title:Prvo destletje restrikcijskega polimorfizma dolžine končnih fragmentov (T-RFLP) v mikrobni ekologiji
Abstract:
Proučevanje restrikcijskega polimorfizma dolžine končnih fragmentov (T-RFLP) tarčnih genov se na področju mikrobne ekologije pričelo šele pred desetletjem. Metoda je hitro postala zelo priljubljena zaradi svoje filogenetske ločljivosti in enostavne analize velikega števila vzorcev. Razvoj priprave vzorcev, zajemanja podatkov, obdelave in standardizacije metod analize so v veliki meri obkrožale silovite razprave. Raziskovalci so tako na vsaki stopnji kritično preizkusili veliko število različnih pristopov. Danes kombinacija tehnik, kot so prag variabilnih deležev, koeficient Bray-Curtis v ne-merskem večdimenzionalnem umerjanju predstavlja ravnotežje med odstranjevanjem suma in zadrževanjem informacij. Tako je nastalo zelo uporabno orodje z relativno enostavno interpretacijo za proučevanje tipizacijskih profilov T-RFLP mikrobnih združb. Njegova trenutna stopnja dovršenosti in predvidene razvojne izpopolnitve v prihodnosti kažejo na velik potencial tega orodja na področju mikrobne ekologije. Hkrati pa bo potrebno uveljaviti tudi bolj standardizirane in bolje kontrolirane izvedbe posameznih stopenj tipizacije mikrobnih združb s T-RFLP.

Keywords:mikrobiologija, mikrobna ekologija, molekularna biologija, molekularna genetika, restrikcijski polimorfizem dolžine končnih fragmentov, T-RFLP

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