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FragmentAnalyser utilities

Usage on BIC/TARS or central-bio

Load the module

To do only once: On central-bio, open a unix shell and if not already done, edit the file .profile (in your home directory) and add this line:

module use /pasteur/projets/policy01/Matrix/modules

quit the shell (or source the .profile file).

When you open a new terminal, you should now be able to use the fragment_analyser module. You can load it in your environment as follows:

module load fragment_analyser

Issue with the creation of images:

echo "backend:agg" >> ~/.config/matplotlib/matplotlibrc

Analyse some files

Assuming you want to analyse a set of files starting with '2015' with csv extensions, type:

fragment_analyser --pattern 2015*csv

If you do not want to create images, add

--no-images

Results are stored in summary_all.csv and summary_filter.csv files.

There are other options related to the analyse. Please type:

fragment_analyser --help

Input files must be CSV with the 12 wells. The number of files is not important.

The tool fragment_analyser will identify the peak across the 12 samples that are the most relevant. All results will be stored in summary_all.csv. If you expect all peaks to be found at the same location, peaks that are outliers should be discarded. In the file summary_filter.csv the outliers are to NA (empty string). Once a peak is identified, we keep track of its position (Size in base pair), concentration and a quantity refered to as nM.

Usage:

fragment_analyser --pattern files*csv

The file results.csv should contains 4 columns:

Well,Sample ID,Size (bp),% (Conc.),nmole/L,ng/ul,Avg. Size,RFU,TIC (ng/ul),TIM (nmole/L),Total Conc. (ng/ ul),amount (nM)
D1,T4,649.0,94.9,11.045,4.3591,716.0,1753.0,4.5912,17.688,4.6007,10.33329382
D2,T7,610.0,91.9,22.672,8.4052,651.0,4773.0,9.1461,44.465,9.1541,21.19848676
D3,P1,667.0,92.7,9.132,3.6994,693.0,1999.0,3.989,20.247,4.0108,8.53281052
D4,Mu,688.0,84.5,3.622,1.5137,678.0,795.0,1.7909,13.168,1.7982,3.384839
D5,rien,,,,,,,,,,
D6,rien,,,,,,,,,,
D7,rien,,,,,,,,,,
D8,rien,,,,,,,,,,
D9,rien,990.0,100.0,0.001,0.0006,982.0,20.0,0.0006,0.001,0.0369,0.0009324
D10,rien,,,,,,,,,,
D11,rien,,,,,,,,,,
D12,Ladder,,,,,,,,,,

The 12th well (Ladder) is always empty.

All columns are copies of input files except for the last column that is the concentration in nM unit. and is computed as:

conc * 1000 / ((Size) * 650 /1000)

where 650 is a hard coded value (mw_dna)

diagnostic.png

More help ?

Please see the notebooks directory or the doc directory. The doc directory is a sphinx project and should be compiled for a better rendering. See also the brief but up-to-date documentation using

fragment_analyser --help