I have a data set of 2x150 reads of 54 pairs of 16S v4 metagenomic sequences from NCBI sra of gastritis patients. When I previously ran the sequences through mothur, the screen.seqs after silva alignment removed sufficient number of sequences.
mothur > screen.seqs(fasta=current, count=current, start=2, end=13426)
Using Ulcer_Donors\stability.trim.contigs.count_table as input file for the count parameter.
Using Ulcer_Donors\stability.trim.contigs.unique.align as input file for the fasta parameter.
Using 4 processors.
It took 1278 secs to screen 3544746 sequences, removed 3505876.
/******************************************/
Running command: remove.seqs(accnos=Ulcer_Donors\stability.trim.contigs.unique.bad.accnos.temp, count=Ulcer_Donors\stability.trim.contigs.count_table)
Removed 5370441 sequences from Ulcer_Donors\stability.trim.contigs.count_table.
Output File Names:
Ulcer_Donors\stability.trim.contigs.pick.count_table
/******************************************/
Output File Names:
Ulcer_Donors\stability.trim.contigs.unique.good.align
Ulcer_Donors\stability.trim.contigs.unique.bad.accnos
Ulcer_Donors\stability.trim.contigs.good.count_table
It took 2067 secs to screen 3544746 sequences.
When I am running the sequences again with the same parameters and commands the result is different. Why is it so?
mothur > screen.seqs(fasta=current, count=current, start=1, end=13424)
Using Ulcer_Donors/stability.trim.contigs.good.count_table as input file for the count parameter.
Using Ulcer_Donors/stability.trim.contigs.good.unique.align as input file for the fasta parameter.
Using 64 processors.
It took 52 secs to screen 2417753 sequences, removed 15177.
/******************************************/
Running command: remove.seqs(accnos=Ulcer_Donors/stability.trim.contigs.good.unique.bad.accnos.temp, count=Ulcer_Donors/stability.trim.contigs.good.count_table)
Removed 18156 sequences from Ulcer_Donors/stability.trim.contigs.good.count_table.
Output File Names:
Ulcer_Donors/stability.trim.contigs.good.pick.count_table
/******************************************/
Output File Names:
Ulcer_Donors/stability.trim.contigs.good.unique.good.align
Ulcer_Donors/stability.trim.contigs.good.unique.bad.accnos
Ulcer_Donors/stability.trim.contigs.good.good.count_table
It took 72 secs to screen 2417753 sequences.
This time I had put in an extra screen.seqs (to put in minlength ) which reduced the number of sequences sufficiently after make.contigs (to put in maxee, deltaq, insert). Could it be because of this?