diff --git a/inst/assets/bibliography.bib b/inst/assets/bibliography.bib index 09b77ff01..dc5e1ef0f 100644 --- a/inst/assets/bibliography.bib +++ b/inst/assets/bibliography.bib @@ -2150,4 +2150,26 @@ @article{Legendre2001 pages = {271--280}, doi = {10.1007/s004420100716}, year = {2001} -} \ No newline at end of file +} + +@article{Schloss2024rarefaction1, + author = {Patrick D. Schloss}, + title = {Rarefaction is currently the best approach to control for uneven sequencing effort in amplicon sequence analyses}, + journal = {{mSphere}}, + volume = {9}, + number = {2}, + pages = {e00354-23}, + doi = {10.1128/msphere.00354-23}, + year = {2024} +} + +@article{Schloss2024rarefaction2, + author = {Patrick D. Schloss}, + title = {Waste not, want not: revisiting the analysis that called into question the practice of rarefaction}, + journal = {{mSphere}}, + volume = {9}, + number = {1}, + pages = {e00355-23}, + doi = {10.1128/msphere.00355-23}, + year = {2024} +} diff --git a/inst/pages/12_quality_control.qmd b/inst/pages/12_quality_control.qmd index 09aa7338a..e60f07846 100644 --- a/inst/pages/12_quality_control.qmd +++ b/inst/pages/12_quality_control.qmd @@ -325,9 +325,10 @@ plotColData(tse,"sum","SampleType", colour_by = "SampleType") + ``` In addition, data can be rarefied with -[subsampleCounts](https://microbiome.github.io/mia/reference/subsampleCounts.html), -which normalises the samples to an equal number of reads. However, -this practice has been discouraged for the analysis of differentially +[rarefyAssay](https://microbiome.github.io/mia/reference/rarefyAssay.html), +which normalises the samples to an equal number of reads. +This remains controversial, however, and strategies to mitigate the information loss in rarefaction have been proposed [@Schloss2024rarefaction1] [@Schloss2024rarefaction2]. +Moreover, this practice has been discouraged for the analysis of differentially abundant microorganisms (see [@mcmurdie2014waste]). ### Contaminant sequences diff --git a/inst/pages/98_exercises.qmd b/inst/pages/98_exercises.qmd index 254c41063..4e56e1cea 100644 --- a/inst/pages/98_exercises.qmd +++ b/inst/pages/98_exercises.qmd @@ -576,7 +576,7 @@ colData(tse)$Barcode_full_length ### Library sizes 1. Calculate library sizes -2. Subsample / rarefy the counts (see: subsampleCounts) +2. Subsample / rarify the counts (see: rarefyAssay) Useful functions: nrow, ncol, dim, summary, table, quantile, unique, scater::addPerCellQC, mia::mergeFeaturesByRank @@ -1296,4 +1296,3 @@ experiments in the MAE. Great job! You performed a cross-correlation analysis between two experiments of a MAE and visualized the results with a heatmap. You are also able to customise the correlation method and significance testing used for the analysis. -