The BiMiCo (Biomap Microbiome Core-tools) package offers single-step preprocessing of 16S microbial marker gene sequencing raw data files for 454 and Illumina platforms.
The current tutorial and example data is aimed at the simple preprocessing of 454-generated fastq data. For more options and details, please see the "BiMiCo_tutorial_LONG" document. (pls contact me for appropriate demo dataset)
- Install CRAN dependencies
install.packages("BiocManager")
install.packages("ggplot2")
- Install Bioconductor dependencies
BiocManager::install("Biobase")
BiocManager::install("Biostrings")
BiocManager::install("phyloseq")
BiocManager::install("dada2")
library(BiMiCo)
library(ggplot2)
\
- fastq files (directory containing raw fastq files intended for analysis exclusively, no other files):
rawfqs <- "./Demo_data/raw_fqs"
- sample phenodata (sample names in rows, sample variables in columns):
phedat <- read.csv("./Demo_data/phenodata_demo.csv", row.names=1)
- if not already contained in sample phenodata, please specify PRIMER TYPE (amplified region):
phedat$primer_type <- "V3-V4"
- taxonomy database to use with DADA2 (default is Greengenes v.13.5):
txset <- "./Demo_data/gg_13_8_train_set_97.fa.gz"
- directory to write quality-filtered fastq files to:
filtered_fqs <- "./Demo_data/filt_fqs"
- directory to write various outputs(ASV table, figures & graphs) to:
out_main <- "./Demo_data/results/"
Generate a summary report based on the desired variable column of phenodata (case/control, sample source, sequencing batch...)
- UNDER DEVELOPMENT: specify phyloseq object, variable of interest, output file basename- PDF of ordination and top 4 ASVs, extended in the future
bimico_summary(toy_set, "clinical_group", "ex1")