From 00ef29fcae324149fc863db06966e2374fc2644f Mon Sep 17 00:00:00 2001
From: AnnaPonomarevaSkoltech
 <123084993+AnnaPonomarevaSkoltech@users.noreply.github.com>
Date: Thu, 26 Jan 2023 16:24:56 +0300
Subject: [PATCH 1/2] Delete README.md

---
 README.md | 2 --
 1 file changed, 2 deletions(-)
 delete mode 100644 README.md

diff --git a/README.md b/README.md
deleted file mode 100644
index 661225e..0000000
--- a/README.md
+++ /dev/null
@@ -1,2 +0,0 @@
-# Final-Project
-Nikita and Oksana are cool

From 1c6bc33241755a61f103570b28b82986c3f58cbb Mon Sep 17 00:00:00 2001
From: AnnaPonomarevaSkoltech
 <123084993+AnnaPonomarevaSkoltech@users.noreply.github.com>
Date: Thu, 26 Jan 2023 16:38:10 +0300
Subject: [PATCH 2/2] Add files via upload

---
 Snakefile Anna | 116 +++++++++++++++++++++++++++++++++++++++++++++++++
 1 file changed, 116 insertions(+)
 create mode 100644 Snakefile Anna

diff --git a/Snakefile Anna b/Snakefile Anna
new file mode 100644
index 0000000..67c7255
--- /dev/null
+++ b/Snakefile Anna	
@@ -0,0 +1,116 @@
+import os
+
+"""
+Snakemake version 6.10.0 <specify your version here>
+Python version 3.10.9 <specify your version here>
+
+To run this code, write in terminal:
+snakemake --use-conda -c <specify number of threads> -n 
+"""
+
+# define samples
+SAMPLES = ['AI-69_S60', 'AI-70_S61', 'AI-71_S62', 'AI-72_S63', 'AI-73_S64']
+REFERENCES = ['T5']
+
+# define directories
+reference_dir = os.path.join('data', 'reference')
+raw_reads_dir = os.path.join('data', 'raw_reads')
+mapped_dir = os.path.join('mapped')
+sorted_dir = os.path.join('mapped_sorted')
+calling_dir = os.path.join('calling')
+result_dir = os.path.join('results')
+
+# create directories
+os.makedirs(reference_dir, exist_ok=True)
+os.makedirs(raw_reads_dir, exist_ok=True)
+os.makedirs(mapped_dir, exist_ok=True)
+os.makedirs(sorted_dir, exist_ok=True)
+os.makedirs(calling_dir, exist_ok=True)
+os.makedirs(result_dir, exist_ok=True)
+
+
+rule all:
+    input:
+        expand(os.path.join(result_dir, 'sorted_{reference}_sample_{sample}_flagstat.log'), reference=REFERENCES, sample=SAMPLES),
+        expand(os.path.join(calling_dir, 'variants_filtered_{reference}_sample_{sample}.vcf'), reference=REFERENCES, sample=SAMPLES)
+
+rule index_reference:
+    input:
+        os.path.join(reference_dir, '{reference}_sequence.fasta')
+    output:
+        os.path.join(reference_dir, '{reference}_sequence.fasta.amb'),
+        os.path.join(reference_dir, '{reference}_sequence.fasta.ann'),
+        os.path.join(reference_dir, '{reference}_sequence.fasta.bwt'),
+        os.path.join(reference_dir, '{reference}_sequence.fasta.pac'),
+        os.path.join(reference_dir, '{reference}_sequence.fasta.sa')
+    conda:
+        os.path.join('envs', 'bwa_env.yml')
+    shell:
+        "bwa index {input}"
+
+rule bwa_map:
+    input:
+        index_amb = os.path.join(reference_dir, '{reference}_sequence.fasta.amb'),
+        index_ann = os.path.join(reference_dir, '{reference}_sequence.fasta.ann'),
+        index_bwt = os.path.join(reference_dir, '{reference}_sequence.fasta.bwt'),
+        index_pac = os.path.join(reference_dir, '{reference}_sequence.fasta.pac'),
+        index_sa = os.path.join(reference_dir, '{reference}_sequence.fasta.sa'),
+        fasta = os.path.join(reference_dir, '{reference}_sequence.fasta'),
+        fastq1 = os.path.join(raw_reads_dir, '{sample}_R1_001.fastq.gz'),
+        fastq2 = os.path.join(raw_reads_dir, '{sample}_R2_001.fastq.gz')
+    output:
+        bam = os.path.join(mapped_dir, '{reference}_sample_{sample}.bam')
+    conda:
+        os.path.join('envs', 'bwa_env.yml')
+    threads: 8
+    shell:
+        "bwa mem -t {threads} {input.fasta} {input.fastq1} {input.fastq2} | samtools view -Sb > {output.bam}"
+
+rule get_statistics:
+    input:
+        os.path.join(sorted_dir, 'sorted_{reference}_sample_{sample}.bam')
+    output:
+        os.path.join(result_dir, 'sorted_{reference}_sample_{sample}_flagstat.log')
+    threads: 1
+    conda:
+        os.path.join('envs', 'samtools_env.yml')
+    shell:
+        "samtools flagstat {input} > {output}"
+
+rule sam_sort:
+    input:
+        os.path.join(mapped_dir, '{reference}_sample_{sample}.bam')
+    output:
+        os.path.join(sorted_dir, 'sorted_{reference}_sample_{sample}.bam')
+    threads: 7
+    conda:
+        os.path.join('envs', 'samtools_env.yml')
+    shell:
+        "samtools sort -@ {threads} {input} -o {output}"
+
+rule sam_index:
+    input:
+        os.path.join(sorted_dir, 'sorted_{reference}_sample_{sample}.bam')
+    output:
+        os.path.join(sorted_dir, 'sorted_{reference}_sample_{sample}.bam.bai')
+    threads: 8
+    conda:
+        os.path.join('envs', 'samtools_env.yml')
+    shell:
+        "samtools index -@ {threads} {input} {output}"
+
+rule variant_calling:
+    input:
+        fasta=os.path.join(reference_dir, '{reference}_sequence.fasta'),
+        bam=os.path.join(sorted_dir, 'sorted_{reference}_sample_{sample}.bam'),
+        index_bai=os.path.join(sorted_dir, 'sorted_{reference}_sample_{sample}.bam.bai')
+    output:
+        os.path.join(calling_dir, 'variants_filtered_{reference}_sample_{sample}.vcf')
+    conda:
+        os.path.join('envs', 'bcftools_env.yml')
+    shell:
+        """
+        bcftools mpileup -Ou -f {input.fasta} {input.bam} | \
+        bcftools call -Ou -mv --ploidy 1 | \
+        bcftools filter -s LowQual > {output}
+        """