From 270c74c522e3a39bcd85831802e81d681dbe92ff Mon Sep 17 00:00:00 2001 From: alvaro Date: Thu, 9 Aug 2018 21:56:23 +0200 Subject: [PATCH 1/8] images used in README.md --- .../normalize/images/effect_nucl_cov.png | Bin 0 -> 234985 bytes user_utils/normalize/images/effect_prot_id.png | Bin 0 -> 9313 bytes .../normalize/images/singleton_len_nucl.png | Bin 0 -> 25028 bytes .../normalize/images/singleton_len_prot.png | Bin 0 -> 26187 bytes 4 files changed, 0 insertions(+), 0 deletions(-) create mode 100644 user_utils/normalize/images/effect_nucl_cov.png create mode 100755 user_utils/normalize/images/effect_prot_id.png create mode 100755 user_utils/normalize/images/singleton_len_nucl.png create mode 100755 user_utils/normalize/images/singleton_len_prot.png diff --git a/user_utils/normalize/images/effect_nucl_cov.png b/user_utils/normalize/images/effect_nucl_cov.png new file mode 100644 index 0000000000000000000000000000000000000000..8641724ff55bf369937fe8206c8d47827d120952 GIT binary patch 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zI6gYM6gPlx9&BfrnL2pHKp1FOd31b?i-%_)E?dPo9O+jri;eJoaXHyxMc?(Vhb@6g z#2I#%FI`=?L`04NeUO!v#hj!9y7LJfW+3Qr5Pk-ZS+L!~TJ*dQxKvS?9}HsdHgL*M zN?J&0A8KPagkg4N}N(_7=rlw9|nywLO-aiDw>JHw7VR zNFeNL=7*%1KMDK-ohYR9wez896c~tb&7^zy?~l-6;qViQME55P6@2=1hm$j;#nkW5 z{uo&KkupmSyB7ByneB<=2mS~o=SN@ypNs=n7aIqMl9Do9B?E}i{Mqvs|8xE_6o&R- gfB8QJ$a5En>f80P)LryQ_!orIL-hwma;7i;3m2xW?*IS* literal 0 HcmV?d00001 From 0f36165a9fefa0289c0dd53a273d020991dfb529 Mon Sep 17 00:00:00 2001 From: alvaro Date: Thu, 9 Aug 2018 22:06:11 +0200 Subject: [PATCH 2/8] second README.md version --- user_utils/normalize/README.md | 55 ++++++++++++++++++++++++++++++++++ 1 file changed, 55 insertions(+) diff --git a/user_utils/normalize/README.md b/user_utils/normalize/README.md index 1d1b9ab..4330ee9 100644 --- a/user_utils/normalize/README.md +++ b/user_utils/normalize/README.md @@ -69,3 +69,58 @@ perl compare_clusters.pl -m -o intersection -d \ your_sequences_homologues/clusters_0taxa_algOMCL_e0_raw,your_sequences_homologues/clusters_0taxa_algOMCL_e0_norm \ &> log.intersection ``` + +## Effect on the clustering + +1) Protein datasets + +Sequence clusters produced by the standard and the normalized version of GET_HOMOLOGUES-EST +on the peptides of 119 isolates of the bacterial Streptophomonas genus showed some differences. +There were 23,249 identical clusters in both runs, but 1,190 and 1,359 clusters unique +to the standard and normalized set up, respectively. The number of clusters containing a single sequence +(singletons) was greater (329 vs 225) and more distributed across peptide length after the normalization step (Figure 3). + +[singleton_dist_prot](images/singleton_len_prot.png) +Figure 3. Length distribution of singletons in the original and normalized clustering. Singleton +sequence length after normalization is more evenly distributed across peptide length. + +Some sequences originally found in clusters were clasified as singletons after normalization. +Subtracting those sequences did not have any effect on the overall percentage of sequence identity of the clusters, +which indicates these may be miss-clustered after normalization. However, some outliers among long sequences +increase cluster mean identity when moved to singleton clusters, indicating the use of the normalization process +for building high quality clusters of long sequences to be used in phylogenetic analyses (Figure 4). + +[diff_identity](images/effect_prot.png) +Figure 4. Difference in cluster % sequence identity before and after removing sequences because of +the normalization process in different length regions. Positive values indicate an increase in identity +after removing a sequence after normalization. Length was measured as the mean alignment length +reported by BLASTP for sequences within the cluster. + +2) Nucleotide datasets + +The clusters sets produced by the standard and normalized predictions by the +GET_HOMOLOGUES-EST protocol were very different with the transcripts of 11 species of the genus Oryza. +In particular, there were 111,964 identical gene clusters, and 14,779 and 38,524 unique clusters in the original +and normalized results, respectively. Moreover, the number of singletons calculated by the standard +program was 612, whereas after normalization it increased to 18,126. The number of singletons +was, as in the protein dataset example, more distributed across nucleotide length after the +normalization step (Figure 5). + +[singleton_dist_nucl](images/singleton_len_nucl.png) +Figure 5. Length distribution of singletons in the original and normalized clustering. Singleton +sequence length after normalization is more evenly distributed across peptide length. + +The mean BLAST coverage values of the clusters usually increased after the subtraction of +sequences because of the normalization. Manual inspection of some cases revealed that long sequences +were subtracted from the original clusters and classified assingletons even when some regions +aligned without mismatches with other sequences of the cluster (Figure 6). This effect of the normalization +process might not be desired if users want to make clusters of CDS and transcripts, even if they only +share some particular regions, such as exons, but not other regions such as introns, only present +in transcripts. In most cases, susbtracting sequences because of normalization did not have an effect on +the overall indentity of the clusters. + +[effect_cov_nucl](images/effect_nucl_cov.png) +Figure 6. Coverage of the clusters before (x-axis) and after (y-axis) new singletons were sunstracted +from original clusters because of the normalization process. + + From 7aaae88ba4cd59e731858f512e8defb197c12b01 Mon Sep 17 00:00:00 2001 From: alvaro Date: Thu, 9 Aug 2018 22:08:05 +0200 Subject: [PATCH 3/8] second README.md version --- user_utils/normalize/README.md | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/user_utils/normalize/README.md b/user_utils/normalize/README.md index 4330ee9..2e29960 100644 --- a/user_utils/normalize/README.md +++ b/user_utils/normalize/README.md @@ -80,7 +80,7 @@ There were 23,249 identical clusters in both runs, but 1,190 and 1,359 clusters to the standard and normalized set up, respectively. The number of clusters containing a single sequence (singletons) was greater (329 vs 225) and more distributed across peptide length after the normalization step (Figure 3). -[singleton_dist_prot](images/singleton_len_prot.png) +![singleton_dist_prot](images/singleton_len_prot.png) Figure 3. Length distribution of singletons in the original and normalized clustering. Singleton sequence length after normalization is more evenly distributed across peptide length. @@ -90,7 +90,7 @@ which indicates these may be miss-clustered after normalization. However, some o increase cluster mean identity when moved to singleton clusters, indicating the use of the normalization process for building high quality clusters of long sequences to be used in phylogenetic analyses (Figure 4). -[diff_identity](images/effect_prot.png) +![diff_identity](images/effect_prot.png) Figure 4. Difference in cluster % sequence identity before and after removing sequences because of the normalization process in different length regions. Positive values indicate an increase in identity after removing a sequence after normalization. Length was measured as the mean alignment length @@ -106,7 +106,7 @@ program was 612, whereas after normalization it increased to 18,126. The number was, as in the protein dataset example, more distributed across nucleotide length after the normalization step (Figure 5). -[singleton_dist_nucl](images/singleton_len_nucl.png) +![singleton_dist_nucl](images/singleton_len_nucl.png) Figure 5. Length distribution of singletons in the original and normalized clustering. Singleton sequence length after normalization is more evenly distributed across peptide length. @@ -119,7 +119,7 @@ share some particular regions, such as exons, but not other regions such as intr in transcripts. In most cases, susbtracting sequences because of normalization did not have an effect on the overall indentity of the clusters. -[effect_cov_nucl](images/effect_nucl_cov.png) +![effect_cov_nucl](images/effect_nucl_cov.png) Figure 6. Coverage of the clusters before (x-axis) and after (y-axis) new singletons were sunstracted from original clusters because of the normalization process. From 0f78238083d0c0bb178ea72e94131bfbf3569000 Mon Sep 17 00:00:00 2001 From: alvaro Date: Thu, 9 Aug 2018 22:09:57 +0200 Subject: [PATCH 4/8] second README.md version --- user_utils/normalize/README.md | 20 ++++++++++++-------- 1 file changed, 12 insertions(+), 8 deletions(-) diff --git a/user_utils/normalize/README.md b/user_utils/normalize/README.md index 2e29960..a875d3e 100644 --- a/user_utils/normalize/README.md +++ b/user_utils/normalize/README.md @@ -78,20 +78,22 @@ Sequence clusters produced by the standard and the normalized version of GET_HOM on the peptides of 119 isolates of the bacterial Streptophomonas genus showed some differences. There were 23,249 identical clusters in both runs, but 1,190 and 1,359 clusters unique to the standard and normalized set up, respectively. The number of clusters containing a single sequence -(singletons) was greater (329 vs 225) and more distributed across peptide length after the normalization step (Figure 3). +(singletons) was greater (329 vs 225) and more distributed across peptide length after the normalization step (Figure 1). ![singleton_dist_prot](images/singleton_len_prot.png) -Figure 3. Length distribution of singletons in the original and normalized clustering. Singleton + +Figure 1. Length distribution of singletons in the original and normalized clustering. Singleton sequence length after normalization is more evenly distributed across peptide length. Some sequences originally found in clusters were clasified as singletons after normalization. Subtracting those sequences did not have any effect on the overall percentage of sequence identity of the clusters, which indicates these may be miss-clustered after normalization. However, some outliers among long sequences increase cluster mean identity when moved to singleton clusters, indicating the use of the normalization process -for building high quality clusters of long sequences to be used in phylogenetic analyses (Figure 4). +for building high quality clusters of long sequences to be used in phylogenetic analyses (Figure 2). ![diff_identity](images/effect_prot.png) -Figure 4. Difference in cluster % sequence identity before and after removing sequences because of + +Figure 2. Difference in cluster % sequence identity before and after removing sequences because of the normalization process in different length regions. Positive values indicate an increase in identity after removing a sequence after normalization. Length was measured as the mean alignment length reported by BLASTP for sequences within the cluster. @@ -104,23 +106,25 @@ In particular, there were 111,964 identical gene clusters, and 14,779 and 38,524 and normalized results, respectively. Moreover, the number of singletons calculated by the standard program was 612, whereas after normalization it increased to 18,126. The number of singletons was, as in the protein dataset example, more distributed across nucleotide length after the -normalization step (Figure 5). +normalization step (Figure 3). ![singleton_dist_nucl](images/singleton_len_nucl.png) -Figure 5. Length distribution of singletons in the original and normalized clustering. Singleton + +Figure 3. Length distribution of singletons in the original and normalized clustering. Singleton sequence length after normalization is more evenly distributed across peptide length. The mean BLAST coverage values of the clusters usually increased after the subtraction of sequences because of the normalization. Manual inspection of some cases revealed that long sequences were subtracted from the original clusters and classified assingletons even when some regions -aligned without mismatches with other sequences of the cluster (Figure 6). This effect of the normalization +aligned without mismatches with other sequences of the cluster (Figure 4). This effect of the normalization process might not be desired if users want to make clusters of CDS and transcripts, even if they only share some particular regions, such as exons, but not other regions such as introns, only present in transcripts. In most cases, susbtracting sequences because of normalization did not have an effect on the overall indentity of the clusters. ![effect_cov_nucl](images/effect_nucl_cov.png) -Figure 6. Coverage of the clusters before (x-axis) and after (y-axis) new singletons were sunstracted + +Figure 4. Coverage of the clusters before (x-axis) and after (y-axis) new singletons were sunstracted from original clusters because of the normalization process. From 63b9b084775207acb714c4cbc8738c674ca0afc8 Mon Sep 17 00:00:00 2001 From: alvaro Date: Thu, 9 Aug 2018 22:12:09 +0200 Subject: [PATCH 5/8] second README.md version --- user_utils/normalize/README.md | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/user_utils/normalize/README.md b/user_utils/normalize/README.md index a875d3e..3cf5c67 100644 --- a/user_utils/normalize/README.md +++ b/user_utils/normalize/README.md @@ -91,12 +91,12 @@ which indicates these may be miss-clustered after normalization. However, some o increase cluster mean identity when moved to singleton clusters, indicating the use of the normalization process for building high quality clusters of long sequences to be used in phylogenetic analyses (Figure 2). -![diff_identity](images/effect_prot.png) +![diff_identity](images/effect_prot_id.png) -Figure 2. Difference in cluster % sequence identity before and after removing sequences because of +_**Figure 2. Difference in cluster % sequence identity before and after removing sequences because of the normalization process in different length regions. Positive values indicate an increase in identity after removing a sequence after normalization. Length was measured as the mean alignment length -reported by BLASTP for sequences within the cluster. +reported by BLASTP for sequences within the cluster.**_ 2) Nucleotide datasets From 7d8f8523448b417466e14c1dc334ababaada570b Mon Sep 17 00:00:00 2001 From: alvaro Date: Thu, 9 Aug 2018 22:13:40 +0200 Subject: [PATCH 6/8] second README.md version --- user_utils/normalize/README.md | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/user_utils/normalize/README.md b/user_utils/normalize/README.md index 3cf5c67..4b6eb39 100644 --- a/user_utils/normalize/README.md +++ b/user_utils/normalize/README.md @@ -82,8 +82,8 @@ to the standard and normalized set up, respectively. The number of clusters cont ![singleton_dist_prot](images/singleton_len_prot.png) -Figure 1. Length distribution of singletons in the original and normalized clustering. Singleton -sequence length after normalization is more evenly distributed across peptide length. +_**Figure 1. Length distribution of singletons in the original and normalized clustering. Singleton +sequence length after normalization is more evenly distributed across peptide length.**_ Some sequences originally found in clusters were clasified as singletons after normalization. Subtracting those sequences did not have any effect on the overall percentage of sequence identity of the clusters, @@ -110,8 +110,8 @@ normalization step (Figure 3). ![singleton_dist_nucl](images/singleton_len_nucl.png) -Figure 3. Length distribution of singletons in the original and normalized clustering. Singleton -sequence length after normalization is more evenly distributed across peptide length. +_**Figure 3. Length distribution of singletons in the original and normalized clustering. Singleton +sequence length after normalization is more evenly distributed across peptide length.**_ The mean BLAST coverage values of the clusters usually increased after the subtraction of sequences because of the normalization. Manual inspection of some cases revealed that long sequences @@ -124,7 +124,7 @@ the overall indentity of the clusters. ![effect_cov_nucl](images/effect_nucl_cov.png) -Figure 4. Coverage of the clusters before (x-axis) and after (y-axis) new singletons were sunstracted -from original clusters because of the normalization process. +_**Figure 4. Coverage of the clusters before (x-axis) and after (y-axis) new singletons were sunstracted +from original clusters because of the normalization process.**_ From b43294c63e98eec48bfb3daf5810aa1e083d9070 Mon Sep 17 00:00:00 2001 From: alvaro Date: Thu, 9 Aug 2018 22:15:16 +0200 Subject: [PATCH 7/8] second README.md version --- user_utils/normalize/README.md | 20 ++++++++++---------- 1 file changed, 10 insertions(+), 10 deletions(-) diff --git a/user_utils/normalize/README.md b/user_utils/normalize/README.md index 4b6eb39..226e111 100644 --- a/user_utils/normalize/README.md +++ b/user_utils/normalize/README.md @@ -72,7 +72,7 @@ perl compare_clusters.pl -m -o intersection -d \ ## Effect on the clustering -1) Protein datasets +_1) Protein datasets_ Sequence clusters produced by the standard and the normalized version of GET_HOMOLOGUES-EST on the peptides of 119 isolates of the bacterial Streptophomonas genus showed some differences. @@ -82,8 +82,8 @@ to the standard and normalized set up, respectively. The number of clusters cont ![singleton_dist_prot](images/singleton_len_prot.png) -_**Figure 1. Length distribution of singletons in the original and normalized clustering. Singleton -sequence length after normalization is more evenly distributed across peptide length.**_ +*Figure 1. Length distribution of singletons in the original and normalized clustering. Singleton +sequence length after normalization is more evenly distributed across peptide length.* Some sequences originally found in clusters were clasified as singletons after normalization. Subtracting those sequences did not have any effect on the overall percentage of sequence identity of the clusters, @@ -93,12 +93,12 @@ for building high quality clusters of long sequences to be used in phylogenetic ![diff_identity](images/effect_prot_id.png) -_**Figure 2. Difference in cluster % sequence identity before and after removing sequences because of +*Figure 2. Difference in cluster % sequence identity before and after removing sequences because of the normalization process in different length regions. Positive values indicate an increase in identity after removing a sequence after normalization. Length was measured as the mean alignment length -reported by BLASTP for sequences within the cluster.**_ +reported by BLASTP for sequences within the cluster.* -2) Nucleotide datasets +_2) Nucleotide datasets_ The clusters sets produced by the standard and normalized predictions by the GET_HOMOLOGUES-EST protocol were very different with the transcripts of 11 species of the genus Oryza. @@ -110,8 +110,8 @@ normalization step (Figure 3). ![singleton_dist_nucl](images/singleton_len_nucl.png) -_**Figure 3. Length distribution of singletons in the original and normalized clustering. Singleton -sequence length after normalization is more evenly distributed across peptide length.**_ +*Figure 3. Length distribution of singletons in the original and normalized clustering. Singleton +sequence length after normalization is more evenly distributed across peptide length.* The mean BLAST coverage values of the clusters usually increased after the subtraction of sequences because of the normalization. Manual inspection of some cases revealed that long sequences @@ -124,7 +124,7 @@ the overall indentity of the clusters. ![effect_cov_nucl](images/effect_nucl_cov.png) -_**Figure 4. Coverage of the clusters before (x-axis) and after (y-axis) new singletons were sunstracted -from original clusters because of the normalization process.**_ +*Figure 4. Coverage of the clusters before (x-axis) and after (y-axis) new singletons were sunstracted +from original clusters because of the normalization process.* From 7f04775fd421391f269662400afa24f0c39ba434 Mon Sep 17 00:00:00 2001 From: alvaro Date: Thu, 9 Aug 2018 22:16:14 +0200 Subject: [PATCH 8/8] second README.md version --- user_utils/normalize/README.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/user_utils/normalize/README.md b/user_utils/normalize/README.md index 226e111..aa9cded 100644 --- a/user_utils/normalize/README.md +++ b/user_utils/normalize/README.md @@ -72,7 +72,7 @@ perl compare_clusters.pl -m -o intersection -d \ ## Effect on the clustering -_1) Protein datasets_ +__1) Protein datasets__ Sequence clusters produced by the standard and the normalized version of GET_HOMOLOGUES-EST on the peptides of 119 isolates of the bacterial Streptophomonas genus showed some differences. @@ -98,7 +98,7 @@ the normalization process in different length regions. Positive values indicate after removing a sequence after normalization. Length was measured as the mean alignment length reported by BLASTP for sequences within the cluster.* -_2) Nucleotide datasets_ +__2) Nucleotide datasets__ The clusters sets produced by the standard and normalized predictions by the GET_HOMOLOGUES-EST protocol were very different with the transcripts of 11 species of the genus Oryza.