Brandon W. King
---------------
-Last updated: Oct 16th, 2006
+Last updated: Oct 18th, 2006
-Updated to Mussagl build: 287? (In process to 424)
+Updated to Mussagl build: (In process to 424)
.. Things to add
* New features / change log
- * Comment out anything isn't implemented yet.
+ * (DONE) Comment out anything isn't implemented yet.
* (DONE) List of features that will be implemented in the future.
* Look into the homology mapping of UCSC.
* Add toggle to genomes.
Features to be Implemented
--------------------------
- * Motif editor supporting more than 10 motifs
- (Status: http://woldlab.caltech.edu/cgi-bin/mussa/ticket/122)
- * Save motifs from Mussagl
- (Status: http://woldlab.caltech.edu/cgi-bin/mussa/ticket/133)
-
For an up-to-date list of features to be implemented visit:
http://woldlab.caltech.edu/cgi-bin/mussa/roadmap
For more information about SMN1 visit `NCBI's OMIM
<http://www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=609682>`_.
+The SMN1 data retrieved in this section can be downloaded from the
+`Mussa Example Data
+<http://woldlab.caltech.edu/cgi-bin/mussa/wiki/ExampleData>`_ page if
+you prefer to skip this section of the manual.
+
+
UCSC Genome Browser Method
--------------------------
example we will retrieve SMN1 through the UCSC genome browser located
at http://genome.ucsc.edu/.
-We have made the SMN1 data available at
-http://woldlab.caltech.edu/cgi-bin/mussa/wiki/ExampleData if you
-prefer to skip this section.
.. image:: images/ucsc_genome_browser_home.png
:alt: UCSC Genome Browser
'demo_w30_t20'. Mussa will create a folder with this name to store
the analysis files in once it has been run.
- 2. Choose a `window size`_. For this demo **choose 30**.
-
- 3. Choose a threshold_... for this demo **choose 20**. See the
+ 2. Choose a threshold_... for this demo **choose 20**. See the
Threshold_ section for more detailed information.
+ 3. Choose a `window size`_. For this demo **choose 30**.
+
+
4. Choose the number of sequences_ you would like. For this demo
**choose 3**.
:alt: Steps 1-4
:align: center
-Now click on the 'Browse' button next to the sequence input box and
-then select /examples/seq/human_mck_pro.fa file. Do the same in the
-next two sequence input boxes selecting mouse_mck_pro.fa and
-rabbit_mck_pro.fa as shown below. Note that you can create annotation
-files using the mussa `Annotation File Format`_ to add annotations to
-your sequence.
+First enter the species name of "Human" in the first "Species" text
+box. Now click on the 'Browse' button next to the sequence input box
+and then select /examples/seq/human_mck_pro.fa file. Do the same in
+the next two sequence input boxes selecting mouse_mck_pro.fa and
+rabbit_mck_pro.fa as shown below. Make sure to give them a species
+name as well. Note that you can create annotation files using the
+mussa `Annotation File Format`_ to add annotations to your sequence.
.. image:: images/define_analysis_step2.png
:alt: Choose sequences
:alt: Mussagl Demo
:align: center
-This analysis is now saved in a directory called **demo_w30_t20** in
-the current working directory. If you close and reopen Mussagl, you
-can reload the saved analysis. See `Load an analysis`_ section below
-for details.
+By default your analysis is NOT saved. If you try to close an analysis
+without saving, you will be prompted with a dialog box asking you if
+you would like to save your analysis. The `Saving`_ section for
+details on saving your analysis. When saving, choose directory and
+give the analysis the name **demo_w30_t20**. If you close and reopen
+Mussagl, you will then be able to load the saved analysis. See `Load
+an analysis`_ section below for details.
Load a mussa parameter file
on creating a .mupa file.
Once you have a .mupa file created, load Mussagl and select the **File >
-Load Mussa Parameters** menu option. Select the .mupa file and click
+Create Analysis from File** menu option. Select the .mupa file and click
open.
.. image:: images/load_mupa_menu.png
~~~~~~~~~~~~~~~~
To load a previously run analysis open Mussagl and select the **File >
-Load Analysis** menu option. Select an analysis **directory** and
+Open Existing Analysis** menu option. Select an analysis **directory** and
click open.
.. image:: images/load_analysis_menu.png
3. Motif_
- 4. `Conservation tracks`_
+ 4. `Red conservation tracks`_
- 5. `Motif Toggle`_
+ 5. `Blue conservation tracks`_
6. `Zoom Factor`_ (Base pairs per pixel)
Each of the black bars represents one of the loaded sequences, in this
case the sequence around the gene 'MCK' in human, mouse, and rabbit.
-FIXME: Should I mention the repeats here?
-
Annotation
~~~~~~~~~~
Annotations can be included on any of the sequences using the `Load a
-mussa parameter file`_ method of loading your sequences. You can
-define annotations by location or using an exact sub-sequence and you
-may also choose any color for display of the annotation; see the
-`Annotation File Format`_ section for details.
-
-Note: Currently there is no way to add annotations using the GUI (only
-via the .mupa file). We plan to add this feature in the future, but it
-likely will not make it into the first release.
+mussa parameter file`_ or `Create a new analysis`_ method of loading
+your sequences. You can define annotations by location or using an
+exact sub-sequence or a FASTA sequence of the section of DNA you wish
+to annotate. See the `Annotation File Format`_ section for details.
Motif
Motif shown in light blue on sequence bar.
The only real difference between an annotation and motif in Mussagl is
-that you can define motifs from within the GUI. See the `Motifs`_
-section for more information.
+that you can define motifs and choose a color from within the GUI. See
+the `Motifs`_ section for more information.
-Conservation tracks
-~~~~~~~~~~~~~~~~~~~
+Red conservation tracks
+~~~~~~~~~~~~~~~~~~~~~~~
.. figure:: images/conservation_tracks.png
:alt: Conservation Tracks
bars.
The **red lines** between the sequence bars represent conservation
-between the sequences and **blue lines** represent **reverse
-complement** conservation. The amount of sequence conservation shown
-will depend on the relatedness of your sequences and the `dynamic
-threshold` you are using. Sequences with lots of repeats will cause
-major slow downs in calculating the matches.
+between the sequences (i.e. not reverse complement matches)
+
+The amount of sequence conservation shown will depend on how much your
+sequences are related and the `dynamic threshold`_ you are using.
-Motif Toggle
-~~~~~~~~~~~~
+Blue conservation tracks
+~~~~~~~~~~~~~~~~~~~~~~~~
-.. image:: images/motif_toggle.png
- :alt: Motif Toggle
+.. figure:: images/conservation_tracks.png
+ :alt: Conservation Tracks
:align: center
+
+ Conservations tracks shown as red and blue lines between sequence
+ bars.
-Toggles motifs on and off. This will not turn on and off annotations.
+**Blue lines** represent **reverse complement** conservation relative
+to the sequence attached to the top of the blue line.
-Note: As of the current build (#200), this feature hasn't been
-implemented.
+The amount of sequence conservation shown will depend on how much your
+sequences are related and the `dynamic threshold`_ you are using.
Zoom Factor
:align: center
You can dynamically change the threshold for how strong a match you
-consider the conservation to be with one of two options:
+consider the conservation to be by changing the value in the dynamic
+threshold box.
- 1. Number of base pair matches out of window size.
-
- 2. Percent base pair conservation.
+The value you enter is the minimum number of base pairs that have to
+be matched in order to be considered conserved. The second number that
+you can't change is the `window size`_ you used when creating the
+experiment. The last number is the percent match.
See the Threshold_ section for more information.
2. Total Size of Sequence
3. Current base pair position
+Note that you can **update the species** text box. Make sure to **save your
+experiment** after making this change by selecting **File > Save
+Analysis** from the menu.
Sequence Scroll Bar
~~~~~~~~~~~~~~~~~~~
useful when you have zoomed in using the `zoom factor`_.
+Saving
+------
+
+Save on Close
+~~~~~~~~~~~~~
+
+When ever you create a new analysis or make a change such as
+adding/editing a motif or changing a species name, an asterisk (*)
+will appear in the title of the window showing that there are changes
+that have not been saved. If you close a Mussa window without saving
+changes, Mussa will ask you if you would like to save the changes that
+have been made.
+
+Save Analysis
+~~~~~~~~~~~~~
+
+After making changes, such as updating species names or adding/editing
+motifs, you can save these changes by selecting the **File > Save
+analysis** menu option or pressing **CTRL + S** (PC) or
+**Apple/Command Key + S** (on Mac).
+
+.. image:: images/save_analysis.png
+ :alt: Save analysis
+ :align: center
+
+Save Analysis As
+~~~~~~~~~~~~~~~~
+
+To save a copy of your analysis to a new location, select the **File >
+Save analysis as** menu option and choose a new location and name for
+your analysis.
+
+.. image:: images/save_analysis_as.png
+ :alt: Save analysis
+ :align: center
+
+Save Motif List
+~~~~~~~~~~~~~~~
+
+See `Save Motifs to File`_ in the `Motifs`_ section.
+
+
+Viewing Multiple Analyses
+-------------------------
+
+Some times it is useful to view more than one analysis at a time. To
+do accomplish this, Mussa allows you to open a new Mussa window by
+selecting the **File > New Mussa Window** menu option.
+
+.. image:: images/new_mussa_window_menu.png
+ :alt: New Mussa Window Menu Option
+ :align: center
+
+A new Mussa window will pop up.
+
+.. figure:: images/new_mussa_window.png
+ :alt: New Mussa Window
+ :align: center
+
+ A new Mussa window on the right, in which I have loaded a second
+ experiment.
+
+Now you can create or load an existing analysis, in this new window,
+as described in the `Create/Load Analysis`_ section.
+
+You can view as many analyses as you can fit on your screen or until
+you run out of available RAM. If you notice a rapid decrease in
+performance and hear lots of noise coming from your hard drive, you
+probably ran out of RAM and are now using virtual memory (i.e. much
+much slower). If this happens, you may need to avoid opening as many
+analyses at one time.
+
+
Annotations / Motifs
--------------------
Save Motifs to File
*******************
-Note: Currently not implemented
+Motifs from the `Motif Dialog`_ can be saved to file for use with
+other analyses. If you just want your motifs to be saved with your
+analysis, see the `save analysis`_ section for details.
+To save a motif list, select **File > Save Motifs** menu option. By
+default, Mussa will append .mtl if you do not provide a file extension
+(valid file extensions: .mtl & .txt).
-Motif Dialog
-************
+.. image:: images/save_motifs.png
+ :alt: Save Motifs
+ :align: center
-**New Features:**
-
-Build 276
- * Allow for toggling individual motifs on and off.
-Build 269
- * Field added for naming motifs.
+Motif Dialog
+************
Mussa has the ability to find lab motifs using the `IUPAC Nucleotide
Code`_ for defining a motif. To define a motif, select **Edit > Edit
:alt: "View > Edit Motifs" Menu
:align: center
-You will see a dialog box appear with a "set motifs" button and 10
-rows for defining motifs and the color that will be displayed on the
-sequence. By default all 10 motifs start off as with white as the
-color. In the image below, I changed the color from white to blue to
-make it easier to see. The first text box is for the motif and the
-second box is for the name of the motif. The check box defines whether
-the motif is displayed or not.
+You will see a dialog box appear with a "apply" button in the bottom
+right and one rows for defining motifs and the color that will be
+displayed on the sequence. When you start adding your first motif, an
+additional row will be added. The check box in the first column
+defines whether the motif is displayed or not. The second column is
+the motif display color. The third column is for the name of your
+motif and finally, the fourth column is motif itself.
.. image:: images/motif_dialog_start.png
:alt: Motif Dialog
:align: center
Now let's make a motif **'AT[C or G]CT'**. Using the `IUPAC Nucleotide
-Code`_, type in **'ATSCT'** into the first box and 'My Motif' for the
-name in the second box as shown below.
+Code`_, type in **'ATSCT'** into the motif field and **'My Motif'** for
+the name in the name field as shown below.
+
+Notice how a second row appeared when you started to add the first
+motif. Every time you add a new motif, a new row will appear allowing
+you to add as many motifs as you need.
.. image:: images/motif_dialog_enter_motif.png
:alt: Enter Motif
:align: center
Now choose a color for your motif by clicking on the colored area to
-the left of the motif. In the image above, you would click on the blue
-square, but by default the squares will be white. Remember to choose a
-color that will show up well with a black bar as the background.
+the left of the name field. Remember to choose a color that will show
+up well with a black bar as the background. A good tool for picking a
+color is the `Colour Contrast Analyser
+<http://juicystudio.com/services/colourcontrast.php>`_ by
+`juicystudio.com <http://juicystudio.com/>`_.
.. image:: images/color_chooser.png
:alt: Color Chooser
:align: center
-Once you have selected the color for your motif, click on the 'set
-motifs' button. Notice that if Mussa finds matches to your motif will
-now show up in the main Mussagl window.
+Once you have selected the color for your motif, click on the
+**'apply'** button. Notice that if Mussa finds matches to your motif
+will now show up in the main Mussa window.
Before Motif:
:alt: Sequence bar after motif
:align: center
+To save your motifs with your analysis, see the `save analysis`_
+section. To save your motifs to a file, see the `save motifs to file`_
+section.
+
View Mussa Alignments
---------------------
:alt: Copy sequence
:align: center
+
Saving to an Image
---------------------------------
projects / mussa.git / blobdiff