dist.shared command problem

Dear Sir,
I am new user to mothur, I got stuck at command dist.shared in MiSeq_SOP. It is showing that i have not provided enough valid groups. Please help me to sort the problem- I am mentioning the programme file which i got from command count.groups.
Thanking you.

mothur > count.groups(shared=stability.an.shared)
mice contains 114769.

Total seqs: 114769.

Output File Names:
stability.an.count.summary


mothur > sub.sample(shared=stability.an.shared, size=2241) Sampling 2241 from each group. 0.03

Output File Names:
stability.an.0.03.subsample.shared


mothur > collect.single(shared=stability.an.shared, calc=chao-invsimpson, freq=100) 0.03

Output File Names:
stability.an.mice.chao
stability.an.mice.invsimpson


mothur > rarefaction.single(shared=stability.an.shared, calc=sobs, freq=100)

Using 8 processors.
0.03

Output File Names:
stability.an.groups.rarefaction


mothur > summary.single(shared=stability.an.shared, calc=nseqs-coverage-sobs-invsimpson, subsample=2441) 0.03

Output File Names:
stability.an.groups.ave-std.summary
stability.an.groups.summary


mothur > heatmap.bin(shared=stability.an.0.03.subsample.shared, scale=log2, numotu=50) 0.03

Output File Names:
stability.an.0.03.subsample.0.03.heatmap.bin.svg


mothur > dist.shared(shared=stability.an.shared, calc=thetayc-jclass, subsample=2241)

Using 8 processors.
You have not provided enough valid groups. I cannot run the command.


With Regards! Dr Ch Mashhood Alam Postdoctoral fellow, Dr Alan Schulmann Group, Plant Genomics Lab, Institute of Biotechnology, University of Helsinki, Finland.

How many samples do you have listed in the *.files file that you use as input to make.contigs? It looks like you have one sample - “mice”. dist.shared won’t do anything if you only have one sample, because you can’t calculate a distance between one sample and itself.

Pat