From 44927033c4643ba74e221fb2793b237237427229 Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?Nicola=C3=AF=20Hoffmann?= Date: Wed, 10 Jul 2024 11:34:43 +0200 Subject: [PATCH] Changes to QC pdf Fixes and changes: - Removed "number of novel genes based on isoform number" heatmap in QC pdf (previously page 9) - Increased font size of all plots in QC pdf - Changed colours and added patterns to "Isoforms/Exons proportion" and "Number of 5' and 3' gene extensions" plots to match colours of other plots (Page 5, 9 and 14 of new pdf) - Added ggpattern to environment dependencies --- README.md | 2 +- assets/metro_map.png | Bin 87579 -> 234035 bytes bin/qc.R | 51 ++++++++++++++++++------------------------- environment.yml | 1 + 4 files changed, 23 insertions(+), 31 deletions(-) diff --git a/README.md b/README.md index dbe54aa..9e48698 100755 --- a/README.md +++ b/README.md @@ -14,7 +14,7 @@ ANNEXA works by using only three parameter files (a reference genome, a referenc 2. Transcriptome reconstruction and quantification with [bambu](https://github.com/GoekeLab/bambu) or [StringTie](https://github.com/gpertea/stringtie). 3. Novel classification with [FEELnc](https://github.com/tderrien/FEELnc). 4. Retrieve information from input annotation and format final gtf with 3 level structure: gene -> transcript -> exon. -5. Predict the CDS of novel protein-coding transcripts with [Transdecoder](https://github.com/TransDecoder/TransDecoder). +5. Predict the CDS of novel protein-coding transcripts with [TransDecoder](https://github.com/TransDecoder/TransDecoder). 6. Filter novel transcripts based on [bambu NDR (Novel Discovery Rates)](https://github.com/GoekeLab/bambu) and/or [TransforKmers TSS validation](https://github.com/IGDRion/transforkmers) to assess fulllength transcripts. 7. Perform a quality control of both the full and filtered extended annotations (see [example](https://github.com/igdrion/ANNEXA/blob/master/examples/results/qc_gtf.pdf)). 8. 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zx(D`~**P#D#BY*Q@?StuJ2Uq_&K~acoTFjcuGrdq!ABfxEX`ZjHE#y-_QHR~+l@ej zh8wUShxa5_I(ld{p;R5^MQS{T5pw}LM*7ju|G+i5E7iXxFWo2uD<7?64vQq$`yTC< zHA@kTMwY&1KCNtB5yHQgSEYie4W>MAlxE@go|yr~PDqn0eEU*DmDlg+6F%Q-lko-j z*v=|@z8=}hP@1xzV3dT4?hYnC4-w76e?2Jw*-$rJ5jk?M;*cwU$upLRZJlDZJWx)f ze4L<}x}KBF-(@@h+DRymY>S8NW7Pm>BjhytU>QV=clpuLp3l4E5`22zEc?XWZ_-;q zn8~e_Gu$9p8{{1yLi*6|z8f&5LoSJVRmPAxpS}|fc{3aB;uzO^$v|Y`{q+MyND{b- z#@cT``83AZIYo)TxqGeJta|u5jKG$FOSKNo-*Gn`v>EZ-W=qkWI9x7^jesbG~qJ3Q^n%tO5UXL3>=pe_Vr3DoP_j=L9_Ha)ZF7JYzI39MA)!fwF(k|_Qz`N`8;JF+# zLou<@fVStTZu4wQS|X(_1EQ(X?|nUaYYR-e4dIX%f~F66c4}djgCq1eqTpi2+f$)z zr|#r69X8j}(N)OTTh)<+x@Vb3e^=H1N5HZ$Z#eK(4aTe?Yt)z|64(45WYQU67iH#2 zEaFn5N-r;?^SC9?QDZ_&5!uRfV7wGS`CJajTd-^IPEnuTQa+_90IeHB4+Wd`B!PAH zf1iqd^JO5izBIDOG>%0m+1@0#6J8omb3{{lGWCq-LpBzE&G#Wcip~BT|HzBVKqI^` z% # Nb isoformes iso = gene %>% mutate(isoformes = ifelse(nb_transcripts == 1, "1", "2+")) %>% - ggplot(aes(x = discovery, fill = isoformes)) + + ggplot(aes(x = discovery, fill = interaction(discovery,gene_biotype), pattern = isoformes)) + ggtitle("Proportion of mono versus multi-isoform genes") + - geom_bar(position = "fill", + geom_bar_pattern(position = "fill", alpha = 0.8, colour = "black") + scale_y_continuous(labels = scales::percent) + facet_grid(~ gene_biotype) + - scale_fill_manual(values = palette) + + scale_fill_manual(values = brew) + + scale_pattern_manual(values = c("1" = "stripe", "2+" = "none")) + + guides(fill = "none", pattern = guide_legend(title = "Isoforms")) + theme(axis.title.x = element_blank()) - # Nombre de gène chaque catégorie count = ggplot(data = gene, aes(x = gene_biotype, fill = paste(gene_biotype, discovery))) + ggtitle("Number of genes") + @@ -170,40 +172,28 @@ gene_counts_sample = gene %>% .x)) ) -# Heatmap Nb_isoforms and samples -gene_tx_samples = gene %>% - filter(discovery == "novel") %$% - as.data.frame(table(.$presents_in_sample, .$nb_transcripts)) %>% - ggplot(aes( - x = Var1, - y = Var2, - fill = log(Freq + 1) - )) + - guides(fill=FALSE) + - ggtitle("Number of novel genes (log) based on isoform\nnumber and number of samples with at least 1 count") + - geom_tile() + - labs(x = "Number of samples with at least 1 count", y = "Number of isoforms", fill = - "log(count)") + - scale_fill_viridis() - # 5'-3' extensions count gene_ext = gene %>% filter(ext_5 > 0 | ext_3 > 0) %>% mutate(ext = ifelse(ext_5 > 0 & ext_3 > 0, "5'-3'", ifelse(ext_5 > 0, "5'", "3'"))) %>% - ggplot(aes(x = gene_biotype, fill = ext)) + + ggplot(aes(x = gene_biotype, fill = gene_biotype, pattern = ext, pattern_angle = ext)) + ggtitle("Number of 5' and 3' gene extensions") + - geom_bar(colour = "black", + geom_bar_pattern(colour = "black", alpha = 0.8, position = position_dodge()) + - scale_fill_manual(values = palette) + + scale_fill_manual(values = brew[c(2,4)]) + + scale_pattern_manual(values = c("stripe","stripe","crosshatch")) + + scale_pattern_angle_manual(values = c(45, -45, 45)) + theme(legend.position = "top") + theme(axis.title.x = element_blank()) + - labs(fill = "Gene extension", y = "Count") + + labs(y = "Count") + + guides(fill = "none", + pattern = guide_legend(title = "Gene extension"), + pattern_angle = guide_legend(title = "Gene extension")) + scale_y_log10() + annotation_logticks(sides = "l") - # 5'-3' extensions distribution labels = c("5'", "3'") names(labels) = c("ext_5", "ext_3") @@ -285,14 +275,16 @@ tx_len = transcript %>% # Mono vs multiexonique tx_ex = transcript %>% mutate(exons = ifelse(nb_exons == 1, "1", "2+")) %>% - ggplot(aes(x = tx_discovery, fill = exons)) + + ggplot(aes(x = tx_discovery, fill = interaction(tx_discovery, transcript_biotype), pattern = exons)) + ggtitle("Proportion of mono versus multi exonic transcripts") + - geom_bar(position = "fill", + geom_bar_pattern(position = "fill", alpha = 0.8, colour = "black") + scale_y_continuous(labels = scales::percent) + facet_grid(~ transcript_biotype) + - scale_fill_manual(values = palette) + + scale_fill_manual(values = brew) + + scale_pattern_manual(values = c("1" = "stripe", "2+" = "none")) + + guides(fill = "none", pattern = guide_legend(title = "Exons")) + theme(axis.title.x = element_blank()) # Count @@ -422,7 +414,6 @@ iso gene_counts gene_validate gene_counts_sample -gene_tx_samples gene_ext gene_ext_dist diff --git a/environment.yml b/environment.yml index 1571b13..9c0cb03 100755 --- a/environment.yml +++ b/environment.yml @@ -14,6 +14,7 @@ dependencies: - conda-forge::r-ggpubr - conda-forge::r-ggridges - conda-forge::r-viridis + - conda-forge::r-ggpattern - conda-forge::procps-ng - conda-forge::mkl<=2024.0