(Terminal-Restriction Fragment Length
is a PCR based genetic fingerprinting
technique for the study of microbial
community structure based on variation
in the 16S rRNA gene. These DNA fragments
are commonly separated using capillary
electrophoresis. One of the primers
of a primer pair is labeled with fluorescent
dye and used to amplify a selected
region of a gene of interest by PCR.
The resulting PCR fragment is digested
with one (or more) restriction endonuclease(s)
and the Terminal Restriction Fragments
(T-RFs) are separated with an automated
DNA analyzer (commonly Beckman
CEQ™ , ABI
PRISM®, or MegaBACE™).
Microbial diversity in a community
can be estimated by analyzing the
number and peak heights of TRF patterns.
GeneMarker software has the ability
to produce highly
sensitive and reproducible results
analysis and Automated Ribosomal Intergenic
Spacer Analysis (ARISA)
by adjusting the AFLP
Analysis default settings. Report
tables may be printed or saved for
import into other T-RFLP analysis
software, such as T-REX.
GeneMarker® software's Large
Fragment Sizing Technology expands
and reduces the cost of these techniques
by affording extended multiplexing
Peak table displayed below a sample
electropherogram showing the option
menu for editing items in the peak
table (clicking the right mouse button
with the cursor on a highlighted cell
will activate the editing menu).
An example print report: Comparison
of electropherograms from four samples.
Review Application Note:
GeneMarker® Software for Terminal-Restriction
Fragment Length Polymorphism (T-RFLP)
Data Analysis (PDF)
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Research use only (RUO)