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Significance status of signature: NA #9
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Hi Maksim, I am very glad to hear that you find Regarding your question, usually TAI profiles should be computed on the entire transcriptome (including all genes) and not for a subset, since this may result in misleading outcomes or interpretations. On the technical side, however, computing TAI values and performing the test stats on ~4000 genes should not be the limiting step. To me the error message looks more like there is a Could you try running: fgig_phyloexp_tf <- na.omit(tf(fgig_phyloexp, function(x) log2(x+1))) Within your analysis steps to see if this makes a difference? Also, when you run the other tests: I hope this helps! Cheers, |
Dear Hajk, |
Dear Maksim, Thank you for your kind feedback and I am glad the issue is resolved. It seems like the I hope this helps. Cheers, |
Dear Dr. Hajk-Georg Drost,
Let me thank you for the awesome and very important myTAI library. In my project, I used myTAI to determine the transcriptome age indices of the life cycle stages of different flatworm species. When considering the complete transcriptomes, no problems arose, and very interesting results were obtained. However we are faced with the NaNs produced warning during PlotSignature when analyzing the “reduced” dataset, including only genes of last common ancestor of the studied species:
Plot signature: ' TAI ' and test statistic: ' FlatLineTest ' running 1000 permutations.
$start.arg
$start.arg$shape
[1] Inf
$start.arg$rate
[1] Inf
$fix.arg
NULL
Significance status of signature: NA
Warning:
1: In dgamma(c(0.000122768450181325, 0.000122768450181325, 0.000122768450181325, :
NaN produced
2: In stats::pgamma(real.var, shape = shape, rate = rate, lower.tail = FALSE) :
NaN produced
Used code:
library(edgeR)
library(myTAI)
library(dplyr)
library(phylotools)
fgig_phylostratr <- read.csv2("../../Phylostratr/Fgigantica/Fgigantica_phylostratr_results.tsv", sep="\t", header = T)
fgig_digenea_ancestor <- get.fasta.name(infile="../../Ancestral_pyHAM/Fgigantica_100aa.Digenea_ancestor.fasta")
fgig_expr <- read.csv2("../../Expr_quant/Fgig_decontaminated_salmon_quant/Fgigantica_100aa_unaveraged_TPMs.tsv", sep="\t", header = T)
fgig_exp_filtered <- subset(fgig_expr, GeneIDs %in% fgig_digenea_ancestor)
fgig_exp_filtered_mut <- mutate_all(fgig_exp_filtered[, 1:length(colnames(fgig_exp_filtered))], function(x) as.numeric(as.character(x)))
fgig_exp_filtered_mut$GeneIDs <- fgig_exp_filtered$GeneIDs
fgig_exp_filtered_mut_mean <- data.frame("GeneIDs"=fgig_exp_filtered_mut$GeneIDs, "Egg"=rowMeans(fgig_exp_filtered_mut[, c("Fgig_egg_rep1", "Fgig_egg_rep2", "Fgig_egg_rep3")]),
"Miracidium"=rowMeans(fgig_exp_filtered_mut[, c("Fgig_miracidium_rep1", "Fgig_miracidium_rep2", "Fgig_miracidium_rep3")]),
"Redia"=rowMeans(fgig_exp_filtered_mut[, c("Fgig_redia_rep1", "Fgig_redia_rep2", "Fgig_redia_rep3")]),
"Cercaria"=rowMeans(fgig_exp_filtered_mut[, c("Fgig_cerc_rep1", "Fgig_cerc_rep2", "Fgig_cerc_rep3")]),
"Metacercaria"=rowMeans(fgig_exp_filtered_mut[, c("Fgig_metacerc_rep1", "Fgig_metacerc_rep2", "Fgig_metacerc_rep3")]),
"Juvenile_42_days"=rowMeans(fgig_exp_filtered_mut[, c("Fgig_juv_42d_rep1", "Fgig_juv_42d_rep2", "Fgig_juv_42d_rep3")]),
"Juvenile_70_days"=rowMeans(fgig_exp_filtered_mut[, c("Fgig_juv_70d_rep1", "Fgig_juv_70d_rep2", "Fgig_juv_70d_rep3")]),
"Adult"=rowMeans(fgig_exp_filtered_mut[, c("Fgig_adult_rep1", "Fgig_adult_rep2", "Fgig_adult_rep3")]))
fgig_phylostratr_filtered <- subset(fgig_phylostratr, qseqid %in% fgig_digenea_ancestor)
colnames(fgig_phylostratr_filtered) <- c("GeneIDs", "MRCA", "Phylostratum", "MRCA_name")
fgig_phylomap <- select(fgig_phylostratr_filtered, "Phylostratum", "GeneIDs")
fgig_phyloexp <- MatchMap(fgig_phylomap, fgig_exp_filtered_mut_mean)
fgig_phyloexp_tf <- tf(fgig_phyloexp, function(x) log2(x+1))
pdf("Fgigantica_logTPMs_TAI.digenea_ancestor.pdf", width = 14)
PlotSignature(ExpressionSet = fgig_phyloexp_tf, measure = "TAI",
TestStatistic = "FlatLineTest", xlab="F.gigantica complex life cycle", ylab="TAI: Digenean ancestor genome model", permutations = 1000)
dev.off()
As a result, on the created plot we see “p_flt = NaN”. I guess is that there is not enough observation count in fgig_phyloexp_tf (3772 observations) to run the tests. Is this possible or is there another explanation or assumption? I would be grateful for any help!
Thanks a lot!
Yours sincerely,
Maksim
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