Biopython gives ValueError: Sequences must all be the same length even though sequences are of the same length
This is the first time I'm doing this. I'm trying to create a phylogenetic tree by making a .phy file from my data.
I have a dataframe:
> ndf =
>ESV trunc
>1 esv1 TACGTAGGTG...
>2 esv2 TACGGAGGGT...
>3 esv3 TACGGGGGG...
>7 esv7 TACGTAGGGT...
I checked the length of the elements of the column "trunc":
length_checker = np.vectorize(len)
arr_len = length_checker(ndf['trunc'])
The resulting arr_len gives the same length (=253) for all the elements.
I saved this dataframe as .phy file, which looks like this:
>23 253
>esv1 TACGTAGGTGGCGAGCGTTATCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCACGGCTCAACCGTGGAGGGTCATTGGAAACTGGGAAACTTGAGTGCAGAAGAGGAAAGCGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGG
>esv2 TACGGAGGGTGCAAGCGTTATCCGGATTCACTGGGTTTAAAGGGTGCGTAGGTGGGTTGGTAAGTCAGTGGTGAAATCTCCGAGCTTAACTTGGAAACTGCCATTGATACTATTAATCTTGAATATTGTGGAGGTTAGCGGAATATGTCATGTAGCGGTGAAATGCTTAGAGATGACATAGAACACCAATTGCGAAGGCAGCTGGCTACACATATATTGACACTGAGGCACGAAAGCGTGGGGATCAAACAGG
>esv3 TACGGGGGGGGCAAGCGTTGTTCGGAATTACTGGGCGTAAAGGGCGCGTAGGCGGCCAGACCAAGTCGAGTGTGAAATTGCAGGGCTTAACTTTGCAGGGTCGCTCGATACTGGTCGGCTAGAGTGTGGAAGAGGGTACTGGAATTCCCGGTGTAGCGGTGAAATGCGTAGATATCGGGAGGAACACCAGCGGCGAAGGCGGGTACCTGGGCCAACACTGACGCTGAGGCGCGAAAGCTAGGGGAGCAAACAG
This is similar to the file used in this tutorial.
However, when I run the command
aln = AlignIO.read('msa.phy', 'phylip')
I get
ValueError: Sequences must all be the same length
I don't know why I'm getting this or how to fix it. Any help is greatly appreciated!
Thanks
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1 answer
The problem is with your msa.phy file. Phylip format has certain specifications.
Your file should look like this:
3 253
esv1 TACGTAGGTGGCGAGCGTTATCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCACGGCTCAACCGTGGAGGGTCATTGGAAACTGGGAAACTTGAGTGCAGAAGAGGAAAGCGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGG
esv2 TACGGAGGGTGCAAGCGTTATCCGGATTCACTGGGTTTAAAGGGTGCGTAGGTGGGTTGGTAAGTCAGTGGTGAAATCTCCGAGCTTAACTTGGAAACTGCCATTGATACTATTAATCTTGAATATTGTGGAGGTTAGCGGAATATGTCATGTAGCGGTGAAATGCTTAGAGATGACATAGAACACCAATTGCGAAGGCAGCTGGCTACACATATATTGACACTGAGGCACGAAAGCGTGGGGATCAAACAGG
esv3 TACGGGGGGGGCAAGCGTTGTTCGGAATTACTGGGCGTAAAGGGCGCGTAGGCGGCCAGACCAAGTCGAGTGTGAAATTGCAGGGCTTAACTTTGCAGGGTCGCTCGATACTGGTCGGCTAGAGTGTGGAAGAGGGTACTGGAATTCCCGGTGTAGCGGTGAAATGCGTAGATATCGGGAGGAACACCAGCGGCGAAGGCGGGTACCTGGGCCAACACTGACGCTGAGGCGCGAAAGCTAGGGGAGCAAACAG
Python code:
from Bio import AlignIO
aln = AlignIO.read('msa.phy', 'phylip')
print(aln)
Output:
SingleLetterAlphabet() alignment with 3 rows and 253 columns
TACGTAGGTGGCGAGCGTTATCCGGAATTATTGGGCGTAAAGCG...AGG esv1
TACGGAGGGTGCAAGCGTTATCCGGATTCACTGGGTTTAAAGGG...AGG esv2
TACGGGGGGGGCAAGCGTTGTTCGGAATTACTGGGCGTAAAGGG...CAG esv
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