The interactive application can be launched in R with the following command:
As an alternative method, an RStudio addin (RStudio version 0.99.878 or higher required) installed with the package can be found in the Addins menu (Figure 1). This button allows the direct launch of the application with a single click.
Figure 1: addin of the app in RStudio (RStudio version >= 0.99.878 required)
The application contains three main panels, as described below.
The first panel includes two elements: a dashboard for selection of trimming and filtering operations, and a menu for selection of the input file/s (Fig. 2).
Figure 2: Panel 1, with each compontent indicated with a number. See explanation for each element below
The “operations menu” (Fig. 2, elements 1 to 8) shows the available operations for file processing:
Remove by N(s): removes sequences with a number of Ns (non identified bases) above a selected threshold value
Remove low complexity sequences: remove sequences with a value of complexity above a threshold value
Remove adapters: removes adapters and partial adapters. Adapter sequences from both ends of single or paired read reads can be selected. Sequences can be reverse-complemented before processing. The program also allows to consider indels and/or anchored adapters.
Filter by average quality: computes the average quality of sequences and removes those with a value below a given threshold
Trim low quality 3’ tails: removes the 3’ tails of sequences that are below a given threshold
Trim 3’ or 5’ by a fixed number: removes a fixed number of bases from the 3’ and/or 5’ ends in the complete set of sequences
Filter sequences by length: removes all the sequences with a number of bases below a threshold value
Remove duplicated sequences: removes duplicated reads, conserving only one copy of each sequence present in the file
The “file selection menu” (Fig. 2, elements 9 to 17) contains options to handle the input file (type of file, file selection), buttons to run, clear and reset the aplication, and the “advanced” submenu:
Single-end reads / paired-end reads: type of input files
“FILE” button: to select an input file
“RUN!” button: to run the program
Output format: to select whether the output file should be compressed (.gz) or not
“CLEAR” button: to clear the configuration of the operations menu that have been selected in the first panel, but keeping the input file(s)
“RESET” button: to restart the application, removing the input file(s) and the selected configurations
Selection notificator: information of the path of the selected file/s
Encoding notificator: information of the input file/s encoding
Advanced options button: to select a custom encoding and set the number of reads included in each chunk for processing, as described below
The “advanced options submenu” (Fig. 3) allows to customize some fine aspects of the trimming and filtering process:
Figure 3: Advanced options submenu
Encoding menu: in addition to the default approach used by the program (auto-detection of file encoding), users can select a standard encoding from a list
Chunk size: the program takes this number of reads at random from the file (default: 1000000), for encoding detection
The second panel (“file operations” panel, Fig. 4) shows the operations that were sucessfuly performed on the input file after running the program.