The Tools for T-RFLP data analysis is provided as an Excel file (Additional file 1: Tools for T-RFLP data analysis.xlsm), which includes the necessary Visual Basic macros. All calculations and storage of data in intermediate and final steps are done in the spreadsheets. This may not be the fastest or most efficient way, but it allows the user fast access to the data and enables easy additional manipulation of the data in a, for many researchers familiar, spreadsheet application.