# Alignments and data¶

You can read in an alignment from a file either with the p4 script with the file name as a command-line argument, or using the read() function within p4. Any alignments that are made are put in var.alignments, a list. (If its a fasta file and the sequences are not all the same length, it is not an alignment, it is a SequenceList, and it is put in var.sequenceLists, another list.) So a lot of p4 interactive sessions might start out like this:

yourShellPromptp4 myData.nex p4> a = var.alignments[0]  and a lot of p4 scripts might start out like this: read('myData.nex') a = var.alignments[0]  Saying read() here reads the file, makes an Alignment object from it, and put that object in var.alignments. Then you give it the name a. Then you can do useful things with a. ## Alignment, Part, and Data classes¶ There are 3 classes. • Alignment. A SequenceList subclass, with a lot of functionality in Python. • Part. A data partition, with the sequence data in the C language, for likelihood calculations. Each alignment has at least one part, maybe more. • Data. All the partitions that you want to use. It is at least one Alignment (but perhaps more), with at least one Part (at least one Part for each Alignment, but perhaps more). A Data object is needed for likelihood calculations. It has a C-language representation of the sequences as a list of partitions. An alignment of phylogenetic data in a file can be read in to make an Alignment object, which is good for simple tasks that you might want to do with alignments. However, if you want to calculate the likelihood of the data, then things get more complicated because then we transfer the alignment into the C-language. A Part object encapsulates an alignment (or an alignment partition) in C. A Data object contains all the Part objects that you want to use, together with the alignments that the parts came from. When you do likelihood calculations, your data might be simple– they might be in just one alignment, with only one data partition. In that case your Data object has only one Alignment object, and one Part object. Or you might have 2 alignments, each of which has a single part each, and so your Data object would have 2 alignments and 2 parts. Or perhaps you have a single alignment which is partitioned into 5 genes, in which case your Data object would have one alignment and 5 parts. The way to get phylogenetic data into p4 is with the ‘read()’ function. When you read in alignments from files, Alignment objects are made and stuffed into var.alignments, a list. So a typical script to do something simple (ie not likelihood) with an alignment might start out by reading in a file, like this: read('myAlignment.nex') a = var.alignments[0]  For likelihood calculations you need a Data object, so a typical script for that might start out like this: read('myAlignment.nex') d = Data()  In either case when you read in the file, an Alignment object is made and deposited in var.alignments. It is there whether or not you give it a name, and whether or not you use it to make a Data object. If you need to partition your alignment, then you might do something like: read("myAlignment.nex") read("mySets.nex") # with the partition info a = var.alignments[0] a.setCharPartition('myCharPartitionName') d = Data() # now with more Parts that if the Alignment had not been partitioned  Part objects are made when you make a Data object. The data that you want to use might be in more than one alignment, and that is perfectly acceptable to p4. In fact, it is essential if you are using more than one datatype (eg the DNA sequence for one gene, analysed simultaneously with the protein sequence for another gene). A MrBayes-style data matrix with more than one datatype is not allowed by p4 (nor is it Nexus compliant). So you might do something like the following to read in two alignments: read('myFirstAlignment.nex') read('mySecondAlignment.nex') d = Data()  P4 analyses data partition-by-partition. Separate alignments necessarily make separate data partitions, but you can also split up alignments into separate partitions–that would be done using Nexus sets blocks. To do this you might do something like: read('myAlignment.nex') a = var.alignments[0] read('myNexusSetsBlock.nex') # Defines cp1 a.setCharPartition('cp1') # Applies cp1 to the alignment d = Data() # now with more Parts that if the Alignment had not been partitioned  In order to get a Data object from one alignment that you want to partition into 2 parts together with one unpartitioned alignment, you might do something like the following: read('myFirstAlignment.nex') a = var.alignments[0] read('myNexusSetsBlock.nex') # defines cp1 a.setCharPartition('cp1') read('mySecondAlignment.nex') d = Data()  The d Data instance now contains 3 Part objects (in d.parts) and 2 Alignment objects (in d.alignments). If you are going to use a Data object together with a tree, eg for ML evaluations, Bayesian analysis, or simulation, you need to attach the Data object to the tree, as: t.data = d  ## Nexus charpartitions¶ This week, p4 understands Nexus charset and charpartition commands within Nexus sets blocks. When a Nexus sets block is read in, its contents are added to var.nexusSets, a NexusSets object which can hold charsets and charpartitions. When you want to use a charpartition on an alignment, you would say something like: a = var.alignments[0] a.setCharPartition(theCharPartitionName)  That attaches a copy of var.nexusSets to the alignment. To un-set a charpartition from an alignment, you can say a.setCharPartition(None). Setting a charPartition does not actually do anything to the sequences in an Alignment, rather a charPartition only comes into play when you make a Data object. When you make a data object from your partitioned alignments, you can check that you are indeed dealing with partitioned data by doing a dump() method on your data object. ## Subsetting alignments¶ By subsetting I mean extracting a part of an alignment into its own new alignment. You can subset alignments based on charsets or mask strings. If you do not have a Nexus sets block, you can still use the Nexus character list format to make a mask on the fly using the func.maskFromNexusCharacterList() function, for example: p4> print func.maskFromNexusCharacterList("1 3-5 7 11", 12, invert=1) 010001011101  ## Nexus standard datatype¶ P4 supports Nexus standard datatype. Something I have been looking into is recoding protein alignments into amino acid groups, for example the 6 Dayhoff groups, for analysis. That uses the p4.alignment.Alignment.recodeDayhoff() method or the p4.alignment.Alignment.recodeProteinIntoGroups() method. Here the amino acid symbols are recoded to the numerals. For example in Dayhoff recoding, since C (Cysteine) is in a group by itself, it is simply recoded as 1, but the amino acids S, T, P, A, and G are all recoded as 2, as they are all in the same Dayhoff group. It is rather like recoding DNA into transversions to decrease the effects of saturation. It is based on the notion that changes within groups might be saturated, biased, and difficult to model, while changes between groups are less problematic. Certainly there is loss of information, but (hopefully) the information that is retained is of higher quality. See Kosiol’s AIS, almost invariant sets. ## (Non-parametric) bootstrapping of data¶ Bootstrapping an alignment is trivially easy, and lots of programs do it, p4 included. It is a method in the Alignment class. However, bootstrapping of partitioned data is also supported by p4. It is a method of the Data class. The alignment and partition structure is maintained in the bootstrapping process. ## Dealing with duplicate sequences¶ There is no point in doing a deep phylogenetic analysis with duplicated sequences. (There may be a reason for popgen level work.) If you have any duplicated sequences, p4 will point them out when you read in your data file. P4 will remove the sequences if you tell it to. But often you want those dupes in your final result - generally a tree. So you will want to remove the duplicated sequences before you do your analysis, and then restore the taxa to the resulting tree after. For example, lets say that you read in your data like this: yourShellPrompt p4 d3.nex


and then you get a long message complaining about duplicated sequences, starting with:

 Alignment from file 'd3.nex'
This alignment has duplicate sequences!
Sequence numbers below are 1-based.
sequence 1 (A) is the same as sequence 3 (C).
sequence 1 (A) is the same as sequence 8 (H).
sequence 2 (B) is the same as sequence 5 (E).
sequence 2 (B) is the same as sequence 7 (G).
...


To remove the duplicated sequences and make a Python dictionary file that will be useful later for restoring those removed taxa, you can do something like this:

read('d3.nex')
a = var.alignments[0]
a.checkForDuplicateSequences(removeDupes=True, makeDict=True)
a.writePhylip(fName='d3_noDupes.phy')


(Above I wrote the subsetted alignment as a phylip file, but that need not be- you could write it in Nexus or fasta format as you choose.)

That makes a dictionary file, by default named p4DupeSeqRenameDict.py. Don’t lose it, because it is needed when you restore the deleted duped taxa to the resulting tree.

It removes all but one of the duplicate sequences, and re-names the remaining sequence with a name like p4Dupe1 or p4Dupe2.

Then you can do your analysis, which will go faster because you don’t have duped sequences. At the end, you will generally have a tree. You can restore the removed taxa to the tree using the dictionary file made above. You would do something like:

read('myTree.nex')
t = var.trees[0]
t.restoreDupeTaxa()
t.writeNexus(fName='myRestoredTree.nex')


The restored taxa will by default have compound names.