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Terminal restriction fragment length polymorphism
Terminal Restriction Fragment Length Polymorphism (TRFLP or sometimes T-RFLP) is a molecular biology technique for profiling of microbial communities based on the position of a restriction site closest to a labelled end of an amplified gene. The method is based on digesting a mixture of PCR amplified variants of a single gene using one or more restriction enzymes and detecting the size of each of the individual resulting terminal fragments using a DNA sequencer. The result is a graph image where the X axis represents the sizes of the fragment and the Y axis represents their fluorescence intensity.
==Background==
TRFLP is one of several molecular methods aimed to generate a fingerprint of an unknown microbial community. Other similar methods include DGGE, TGGE, ARISA, ARDRA, PLFA, etc.
These relatively high throughput methods were developed in order to reduce the cost and effort in analyzing microbial communities using a clone library. The method was first described by Liu and colleagues in 1997〔Liu, W, Marsh, T, Cheng, H, & Forney, L (1997) Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA. Appl. Environ. Microbiol. 63: 4516-4522〕 which employed the amplification of the 16S rDNA target gene from the DNA of several isolated bacteria as well as environmental samples.
Since then the method has been applied for the use of other marker genes such as the functional marker gene pmoA to analyze methanotrophic communities.
抄文引用元・出典: フリー百科事典『 ウィキペディア（Wikipedia）』
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