Difference between revisions of "Summary.seqs"

Latest revision as of 17:29, 29 April 2019

The summary.seqs command will summarize the quality of sequences in an unaligned or aligned fasta-formatted sequence file.

Default Setting

Fasta is the only required parameter:

mothur > summary.seqs(fasta=amazon.fasta)


		Start	End	NBases	Ambigs	Polymer	NumSeqs
Minimum:	1	422	422	0	4	1
2.5%-tile:	1	436	436	0	4	3
25%-tile:	1	507	507	1	5	25
Median: 	1	530	530	3	5	50
75%-tile:	1	961	961	6	6	74
97.5%-tile:	1	973	973	15	8	96
Maximum:	1	978	978	20	9	98
Mean:	1	678.235	678.235	4.54082	5.44898
# of Seqs:	98

For this unaligned fasta file, we see that all of the sequences started at position 1 (they're unaligned) and had between 422 and 978 bases in them. The median length was 530 bases. We can also see that more than 75% of the sequences had at least one ambiguous base in them and at least one had 20. The final column indicates the length of the longest homopolymer in each sequence - 95% of the sequences had a homopolymer length between 4 and 8 bases long.

An output file (e.g. amazon.fasta.summary) lists each of these parameters for each sequence. For example:

seqname	start	end	nbases	ambigs	polymer numSeqs
U68589	1	943	943	10	5     1
U68590	1	497	497	0	6     1
U68591	1	930	930	1	4     1
...

So we see that sequence U68589 was 943 bases long, had 10 ambiguous bases in it and the longest homopolymer in the sequence was 5 bases long. The numSeqs column is 1 because no name file was provided so we assume this sequence is unique.

If we had instead analyzed an aligned sequence file such as the greengenes alignment database:

mothur > summary.seqs(fasta=core_set_aligned.imputed.fasta)

		Start	End	NBases	Ambigs	Polymer	NumSeqs
Minimum:	69	6849	1423	0	4	1
2.5%-tile:	97	6849	1469	0	5	124
25%-tile:	109	6849	1507	0	5	1235
Median: 	109	6849	1524	0	6	2470
75%-tile:	109	6849	1538	0	6	3704
97.5%-tile:	109	6857	1563	3	8	4815
Maximum:	109	6885	1609	30	11	4938
Mean:	107.857	6849.47	1521.43	0.222155	5.67294
# of Seqs:	4938

Now we see that all of the sequences are at least 1,423 bases long, very few have any ambiguous bases and most sequences start by position 109 and end by position 6,849. These data can be useful for removing sequences that don't overlap or that have features indicating poor quality using the screen.seqs command.

name

You can also use a name file with the summary.seqs command.

mothur > summary.seqs(fasta=stool.unique.fasta)

		Start	End	NBases	Ambigs	Polymer	NumSeqs
Minimum:	1	183	183	0	3	1
2.5%-tile:	1	242	242	0	4	507
25%-tile:	1	258	258	0	5	5061
Median: 	1	267	267	0	5	10121
75%-tile:	1	274	274	0	5	15181
97.5%-tile:	1	287	287	0	6	19735
Maximum:	1	373	373	0	6	20241
Mean:	1	265.647	265.647	0	4.9541
# of Seqs:	20241

or

mothur > summary.seqs(fasta=stool.unique.fasta, name=stool.names)

		Start	End	NBases	Ambigs	Polymer	NumSeqs
Minimum:	1	183	183	0	3	1
2.5%-tile:	1	243	243	0	4	929
25%-tile:	1	259	259	0	5	9282
Median: 	1	267	267	0	5	18564
75%-tile:	1	274	274	0	5	27845
97.5%-tile:	1	287	287	0	6	36198
Maximum:	1	373	373	0	6	37126
Mean:	1	265.768	265.768	0	4.96205
# of unique seqs:	20241
total # of seqs:	37126

count

The count file is similar to the name file in that it is used to represent the number of duplicate sequences for a given representative sequence.

mothur > summary.seqs(fasta=stool.unique.fasta, count=stool.count_table)

summary && contigsreport && alignreport

The summary parameter allows you to provide a *.summary file as an input file.

mothur > summary.seqs(summary=stability.trim.contigs.good.unique.good.summary)

		Start	End	NBases	Ambigs	Polymer	NumSeqs
Minimum:	1250	11550	250	0	3	1
2.5%-tile:	1968	11550	252	0	4	4711
25%-tile:	1968	11550	252	0	4	47109
Median: 	1968	11550	252	0	4	94218
75%-tile:	1968	11550	253	0	5	141326
97.5%-tile:	1968	11550	253	0	6	183724
Maximum:	1968	13383	259	0	8	188434
Mean:	1967	11550	252	0	4
# of unique seqs:	20336
total # of seqs:	188434

The contigsreport parameter allows you to provide the *contigs.report file as input.

mothur > summary.seqs(contigsreport=stability.contigs.report)

		Length	Overlap_Length	Overlap_Start	Overlap_End	MisMatches	Num_Ns	NumSeqs
Minimum:	246	0	0	83	0	0	1
2.5%-tile:	252	248	1	251	0	0	5699
25%-tile:	252	249	1	251	0	0	56989
Median: 	252	249	2	251	2	0	113978
75%-tile:	253	250	2	251	7	0	170967
97.5%-tile:	253	250	3	252	31	7	222257
Maximum:	501	254	251	263	124	238	227955
Mean:	252	248	2	251	5	0
# of Seqs:	227955

The align report parameter allows you to provide an align.report file as input.

mothur > summary.seqs(alignreport=stability.MISeq_SOP.trim.contigs.good.unique.align.report)

		Length	SimBtwnQueryTemplate	LongestInsert	SearchScore	NumSeqs
Minimum:	250	70	0	12	1
2.5%-tile:	252	86	0	48	411
25%-tile:	252	92	0	63	4107
Median: 	253	94	0	71	8214
75%-tile:	253	97	0	87	12320
97.5%-tile:	254	99	0	96	16016
Maximum:	270	100	3	100	16426
Mean:	252	94	0	74
# of Seqs:	16426

processors

The processors option enables you to accelerate the summary process by using multiple processors. Default processors=Autodetect number of available processors and use all available. You can use 2 processors with the following option:

mothur > summary.seqs(fasta=stool.unique.fasta, name=stool.names, processors=2)

Revisions

• 1.22.0 - Added processors option for Windows users.
• 1.22.0 - Added NumSeqs and mean values to output.
• 1.28.0 - Added count option
• 1.31.0 - Bug Fix: 32bit machines processing a file larger than 4G could hang
• 1.39.1 - Adds file mismatch check
• 1.40.0 - Adds summary, contigsreport, alignreport parameters. #319
• 1.40.0 Rewrite of threaded code. Default processors=Autodetect number of available processors and use all available.
• 1.42.0 Removes extra space from output file names. #603