Tools for T-RFLP data analysis using Excel

Area of Science:

• Microbiology
• Bioinformatics
• Molecular Ecology

Background:

• Terminal Restriction Fragment Length Polymorphism (T-RFLP) is a DNA-fingerprinting technique for microbial community composition analysis.
• Lack of standardized methods for T-RFLP data analysis hinders comparisons across studies.
• Manual analysis of large T-RFLP datasets is impractical; existing automated tools lack flexibility.

Purpose of the Study:

• To develop a flexible analysis template for T-RFLP data using Microsoft Excel.
• To enable objective and efficient analysis of large T-RFLP datasets.
• To facilitate comparisons and combinations of different data treatment methods.

Main Methods:

• Development of a Microsoft Excel-based template with macros for T-RFLP data analysis.
• Implementation of procedures for noise baseline thresholding, analysis range setting, and profile normalization/alignment.
• Inclusion of options for generating consensus profiles and calculating association coefficients and diversity indices.

Main Results:

• The template provides comprehensive procedures for analyzing large T-RFLP datasets.
• Users can customize analysis parameters, including noise baseline, alignment, and normalization methods.
• Calculations can be based on either peak height or peak area data.

Conclusions:

• The developed template offers an objective and flexible approach to T-RFLP data analysis.
• It enables efficient handling of large datasets within a widely accessible spreadsheet application.
• Facilitates robust comparisons of microbial community structures.