TAREAN output description
Table of Contents
1 Introduction
TAREAN output includes HTML report with list of all analyzed clusters; the clusters are classified into five categories:
- high confidence satellites
- low confidence satellites
- potential LTR elements
- rDNA
- other clusters
Each cluster for which consensus sequences was reconstructed has also its own detailed report, linked to the main report.
2 Main HTML report
This report contains basic information about all clusters larger than specified threshold (default value is 0.01% of analyzed reads)
2.1 Table legend
- Cluster
- Cluster identifier
- Genome Proportion
[%] - (Number of sequences in cluster/Number of sequences in clustering) x 100%
- Size
- Number of reads in the cluster
- Satellite probability
- Empirical probability estimate that cluster sequences are derived from satellite repeat. This estimate is based on analysis of more than xxx clusters including yyy manually anotated and zzz experimentaly validated satellite repeats
- Consensus
- Consensus sequence is outcome of kmer-based analysis and represents the most probable satellite monomer sequence
- Kmer analysis
- link to analysis report for individual clusters
- Graph layout
- Graph-based visualization of similarities among sequence reads
- Connected component index
- Proportion of nodes of the graph which are part of the the largest strongly connected component
- Pair completeness index
- Proportion of reads with available mate-pair within the same cluster
- Kmer coverage
- Sum of relative frequencies of all kmers used for consensus sequence reconstruction
- |V|
- Number of vertices of the graph
- |E|
- Number of edges of the graph
- PBS score
- Primer binding site detection score
- The longest ORF length
- Length of the longest open reading frame found in any of the possible six reading frames. Search was done on dimer of consensus so ORFs can be longer than 'monomer' length
- Similarity-based annotation
- Annotation based on similarity search using blastn/blastx against database of known repeats.
3 Detailed cluster report
Cluster report includes a list of major monomer sequence varinats reconstructed from the most frequent k-mers. The reconstructed consensus sequences are sorted based on their significance (that is, what proportion of k-mer they represent).
3.1 Table legend
- kmer
- length of kmer used for consensus reconstruction.
- variant
- identifier of consensus variant.
- total score
- measure of significance of consensus variant. Score is calculated as a sum of weights of all k-mers used for consensus reconstruction.
- monomer length
- length of the consensus
- consensus
- consensus sequence without ambiguous bases.
- graph image
- part of de-Bruijn graph based on the abundant k-mers. Size of vertices corresponds to k-mer frequencies, Paths in the graph which was used for reconstruction of consensus sequences is gray colored.
- logo image
- consensus sequences shown as DNA logo. Height of letters corresponds to kmer frequencies. Logo images are linked to corresponding position probability matrices.
4 Structure of the output archive
Complete results from TAREAN analysis can by downloaded as zip archive which contains the following files and directories:
. . ├── clusters_info.csv <------------ list of clusters in tab delimited format ├── index.html <------------ main html report ├── seqclust │ ├── assembly # not implemented yet │ ├── blastn <------------ results of read comparison with DNA database │ ├── blastx <------------ results of read comparison with protein database │ ├── clustering │ │ ├── clusters │ │ │ ├── dir_CL0001 <----┐- detailed information about clusters │ │ │ ├── dir_CL0002 <----│ │ │ │ ├── dir_CL0003 <----│ │ │ │ .... <----┘ │ │ │ │ │ └── hitsort.cls <--------- list of reads in individual clusters │ ├── mgblast │ ├── prerun │ └── sequences <--------- input reads ├── summary # not implemented yet ├── TR_consensus_rank_1_.fasta <-- reconstructed monomer sequences for HIGH confidence satellites ├── TR_consensus_rank_2_.fasta <-- reconstructed monomer sequences for LOW confidence satellites ├── TR_consensus_rank_3_.fasta <-- reconstructed sequences of potential LTR elements └── TR_consensus_rank_4_.fasta <-- reconstructed consensus for rDNA
List of all clusters which is available in HTML file index.html is also
available in tab delimited format in the file clusters_info.csv which can be
easily viewed and edited in spreadsheet editing programs. List of all clusters
and the corresponding reads is in the file hitsort.cls which has the following
format:
>CL1 11 134234r 55494f 85525f 136746r 96742f 91926f 239729r 105445f 222518r 136402r 9013 >CL2 10 76205r 120735r 69527r 12235r 176778f 189307f 131952f 163507f 100038r 178475r >CL3 6 99835r 222598f 29715r 102023f 99524r 30116f >CL4 6 51723r 69073r 218774r 146425f 136314r 41744f >CL5 5 70686f 65565f 234078r 50430r 68247r
where CL1 11 is the cluster ID followed by number of reads in the cluster;
next line contains list of all read names belonging to the cluster.
4.1 structure of cluster directories
Detailed information for each cluster is stored is subdirectories:
dir_CL0011
├── blast.csv <------------tab delimited file, all-to-all comparison od reads within cluster
├── CL11_directed_graph.RData <----directed graph representation of cluster saved as R igraph object
├── CL11.GL <-----------------undirected graph representation of cluster saved as R igraph object
├── CL11.png <-----------┐- images with graph visualization
├── CL11_tmb.png <-----------┘
├── dna_database_annotation.csv <-- annotation of cluster reads based on the DNA database of repeats
├── reads_all.fas <---------------- all reads included in the cluster in fasta format
├── reads.fas <---------------- subset of reads used for monomer reconstruction
├── reads_oriented.fas <------------ subset of reads all in the same orientation
└── tarean
├── consensus.fasta <----------- fasta file with tandem repeat consensus variants
├── ggmin.RData
├── img
│ ├── graph_11mer_1.png <-----┐
│ ├── graph_11mer_2.png <-----│
│ ├── graph_15mer_2.png <-----│
│ ├── graph_15mer_3.png <-----│
│ ├── graph_15mer_4.png <-----│ images of kmer-based graphs used for reconstruction of
│ ├── graph_19mer_2.png <-----│ monomer variants
│ ├── graph_19mer_4.png <-----│
│ ├── graph_19mer_5.png <-----│
│ ├── graph_23mer_2.png <-----│
│ ├── graph_27mer_3.png <-----┘
│ │
│ ├── logo_11mer_1.png <-----┐
│ ├── logo_11mer_2.png <-----│
│ ├── logo_15mer_2.png <-----│
│ ├── logo_15mer_3.png <-----│
│ ├── logo_15mer_4.png <-----│ images with DNA logos representing consensus sequences
│ ├── logo_19mer_2.png <-----│ of monomer variants
│ ├── logo_19mer_4.png <-----│
│ ├── logo_19mer_5.png <-----│
│ ├── logo_23mer_2.png <-----│
│ └── logo_27mer_3.png <-----┘
│
├── ppm_11mer_1.csv <-----┐
├── ppm_11mer_2.csv <-----│
├── ppm_15mer_2.csv <-----│
├── ppm_15mer_3.csv <-----│
├── ppm_15mer_4.csv <-----│ position probability matrices for individual monomer
├── ppm_19mer_2.csv <-----│ variants derived from k-mer frequencies
├── ppm_19mer_4.csv <-----│
├── ppm_19mer_5.csv <-----│
├── ppm_23mer_2.csv <-----│
├── ppm_27mer_3.csv <-----┘
│
├── reads_oriented.fas_11.kmers <-----┐
├── reads_oriented.fas_15.kmers <-----│
├── reads_oriented.fas_19.kmers <-----│ k-mer frequencies calculated on oriented reads
├── reads_oriented.fas_23.kmers <-----│ for k-mer lengths 11 - 27
├── reads_oriented.fas_27.kmers <-----┘
├── reads_oriented.fasblast_out.cvs <---------┐results of blastn search against database of tRNA
├── reads_oriented.fasblast_out.cvs_L.csv <----│for purposes of LTR detection
├── reads_oriented.fasblast_out.cvs_R.csv <----┘
└── report.html <--- cluster analysisHTML summary