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A set of files to do genomics analysis in python
To use any of these script collections, just run these two lines in your python kernel / Jupyter notebook:
sys.path.append('/home/ubuntu/tools/python-genomics')
import Scanpyplus
He, Lim and Sun et al. A human fetal lung cell atlas uncovers proximal-distal gradients of differentiation and key regulators of epithelial fates
Among the functions in Scanpyplus, there's also a function to do feature gene selection (DeepTree algorithm). It removes garbage among highly variable genes, mitigate batch effect if you remove garbage batch by batch, and increases signal-to-noise ratio of the top PCs to promote rare cell type discovery.
Here is a notebook to use DeepTree algorithm to "de-noise" highly-variable genes and improve initial clustering.
A MATLAB implementation can be found here.
This algorithm can be potentially used to reduce batch effect when fearing overcorrection, especially comparing conditions or time points. Two notebooks are provided showing "soft integration" of fetal limb and pancreas data.
There are 4 types of doublets:
Cross-sample doublets can usually be identified by hastags or genetic backgrounds. Theoretically, (n-1)/n of doublets can be identified as cross-sample doublets when n samples with different hashtags/genetics are pooled equally.
Heterotypic doublets can sometimes trick data scientists into thinking they are a new type of dual-feature cell type like NKT cells etc. Heterotypic doublets are usually identified by matching individual cells to synthetic doublets regardless of manually curated clusters. Algorithms like Scrublet can remove a substantial part of doublet cells but not all of them. The survivor doublets can still aggregate into tiny clusters picked up by the annotaters when doing subclustering. Doublets of rarer cell types are also often missed, which obscures the discoveries of new cell types and states. To leverage the input from biologists' manual parsing and the increased sensitivity of cluster-average signatures, I introduce here an alternative approach to facilitate heterotypic doublet cluster identification. This approach scans through individual tiny clusters and look for its "Parent 2" that gives it a unique feature that's different from its sibling subclusters sharing the same "Parent 1". A notebook using published PBMC data is provided.
Other functions in Scanpyplus:
An alternative way to call doublet subclusters based on Scrublet and the gastrulation paper
Bertie(adata,Resln=1,batch_key='batch') was written with the help from K. Polanski. This script aggregates Scrublet scores from subclusters and makes threshold cuts based on subcluster p-values. And this is done batch by batch.
A variant version Bertie_preclustered allows users to use user-defined clusters to calculate p-values. This is also done batch by batch.
You can extract the color dict of a variable from an anndata object using ExtractColor(adata,obsKey='louvain',keytype=int),
and manipulate the color dict using UpdateUnsColor.
You can also cherry pick a value of a variable and make it white using MakeWhite.
You can plot sankey graph between two variables of an anndata object using ScanpySankey.
Re-ordering the cluster IDs based on relationship rather than size can be done by orderGroups.
remove_barcode_suffix removes the suffix after the '-' in the cell (barcode) name.
CopyMeta copies the metadata (both obs and var) from one object to another.
AddMeta stores a dataframe of obs values per each cell into an object.
AddMetaBatch reads a dataframe of obs values per batch into an object. This format of metadata (rows are batch names, columns are obs categories) is more common, compact and human readable that is usually stored in Excel spreadsheets.
OrthoTranslate translates mouse genes to human orthologs and filter out poorly conserved genes, based on ortholog table that can be derived from Biomart etc.
file2gz creates .gz files which is useful for creating artificial 10X files.
Scanpy2MM saves an anndata into MatrixMarket form.
mtx2df reads MatrixMarket files into a dataframe.
Transfer the raw layer to the default layer by GetRaw and calculate integer raw counts based on n_counts
and log-transformed counts using CalculateRaw.
For large matrices, cells can be DownSampled based on labels such as cell types.
Sometimes PseudoBulk profiles are also useful to generate, whether it's the mean, median or max.
ShiftEmbedding creates a platter that juxtaposes subsets of the data (batches, stages etc.) to visualize side by side.
CopyEmbedding copies the embedding of one object to another.
celltype_per_stage_plot and stage_per_celltype_plot plot horizontal and vertical bar plots respectively based on two metadata variables (cell type and stage, for example).
Plot3DimUMAP generates a 3D plot (by plotly) of the UMAP after sc.tl.umap produces the 3D coordinates.
DEmarkers calculates, filteres and plots differentially expressed genes between two populations.
GlobalMarkers calculates marker genes for every cell cluster and filters them.
ClusterGenes transposes a log-transformed adata object and performs clustering and dimension reduction to classify genes.
Dotplot2D plots the expression levels of a gene across two metadata categories (e.g. samples and cell types). It can be used to trace maternal contimation by plotting XIST and check a key gene's expression patterns against cell types and age etc.
snsSplitViolin plots splitviolin plots for two populations.
snsCluster plots clustermaps using an anndata object as input. This has been helped by Bao Zhang from Zhang lab
markSeaborn marks specific genes on a Seaborn plot.
extractSeabornRows extracts the rowlabels of a Seaborn object and saves into a Series.
Venn_Upset can be used to directly plot upset plots (bar plots of each category of intersections).
LogisticRegressionCellType can learn the defining features of a variable (such as cell type) of the reference object and predict the corresponding labels of a query object.
The saved model files and also be re-used to predict a new query object in future by LogisticPrediction.
Functions in pandasPlus:
DF2Ann converts a dataframe into an anndata object.
UpSetFromLists plots an upset plot (barplot of Venn diagram intersections) based on lists of lists.
show_graph_with_labels plots an interaction graph using edges to represent connection strength (max at 1, at least 0.9 to be shown).
Dataframe values can also be used to calculate zscore and Ginni coefficients.
cellphonedb_n_interaction_Mat and cellphonedb_mat_per_interaction are useful to reformat cellphonedb outputs.