Quick Start Guide

  1. Enter primer set or probe to check.  Enter a primer set or probe  in the text boxes provided within the main window.  Make sure that all oligonucleotides entered correspond to the sense strand of the target nucleic acid, and that they run from 5'-3'.  Use ReverseComplement to convert antisense oligonucleotides, if necessary.  You may also enter positional information relative to the Esherichia coli 16S rRNA gene, if that gene is being considered and the RDP database is to be loaded (this ensures that only sequence records of sufficient length within the RDP database are considered).
  2. Load one or more datasets to check primer set or probe against.  Click 'Load Database(s)' button within the main window and use the resulting dialog box to select  one or more datasets that your primer set or probe will be checked against.  
  3. Run the analysis.   Press the 'Do Analysis' button on the main window - the duration of the resulting analysis will depend mainly on the size of the database loaded and the speed of your computer.  For example, ~12,000 records will take approximately 15 minutes to run on a computer with a 900 MHz processor.
  4. View output.  Press 'View Results' on the main window and a results window is presented.  The databases previously selected are redisplayed, this time colour coded according to how well their contents are described by the primer set or probe used.  Identify the database representing the target group you are interested in (by clicking on it with the mouse) and a graph is displayed showing what percentage of records within and outside of that group are targeted by the oligo(s) with increasing number of mismatches.