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Essential AWK Commands for Next Generation Sequence Analysis

Here are the few essential awk command line scripts for next generation sequence analysis.

Users need latest version of gawk to run commands with bitwise operations. Most of the Linux distributions come with gawk. However OSX users have to install it from here

http://rudix.org/packages/gawk.html

Count number of reads in a FastQ file

awk ‘END{print NR/4}’ file.fastq

Convert FastQ to FastA

 awk 'NR%4==1{a=substr($0,2);}NR%4==2{print ">"a"\n"$0}’ file.fastq

 

Get reads matching a sequence pattern and convert them to FastA

awk 'NR%4==1{a=substr($0,2);}NR%4==2 && $1~/GAATTC/ {print ">"a"\n"$0}’ file.fastq

This will get all reads with EcoRI cleavage (GAATTC)

Separate reads based on their length

awk 'NR%4==1{a=$0} NR%4==2{b=$0} NR%4==3{c=$0} NR%4==0&&length(b)>=75{print a”\n”b”\n”c”\n”$0;}' file.fq

This will print reads thats are 75bp or more in length.

Printing the above output in FastA format

awk 'NR%4==1{a=substr($0,2);} NR%4==2{b=$0} NR%4==3{c=$0} NR%4==0&&length(b)>=75{print a”\n”b”\n”c”\n”$0;}' file.fq

 

Get All header lines from a SAM file

awk '$1~/^@/' file.sam

 

Get all reads – excluding headers

awk '$1!~/^@/' file.sam

 

Get all unmapped 

gawk -F'\t' $1!~/^@/ && and($2,0x4)' file.sam

Get all mapped

gawk -F'\t' '$1!~/^@/ && !and($2,0x4)' file.sam

 

Count unmapped

awk -F'\t' '$1!~/^@/ && and($2,0x4){i++};END{print i}’ file.sam

Count mapped

awk -F'\t' '$1!~/^@/ && !and($2,0x4){i++};END{print i}’ file.sam

 

Convert all unmapped reads into fasta format

awk -F'\t' '$1!~/^@/ && and($2,0x4) {print ">" $1; if(and($2,0x10)) system("echo " $10 "| rev| tr '[ATGC]' '[TACG]'"); else print $10}' file.sam

Convert all mapped reads into fasta format

awk -F'\t' '$1!~/^@/ && !and($2,0x4) {print ">" $1; if(and($2,0x10)) system("echo " $10 "| rev| tr '[ATGC]' '[TACG]'"); else print $10}' file.sam

Convert all reads into fasta format

awk -F'\t' '$1!~/^@/{print ">" $1; if(and($2,0x10)) system("echo " $10 "| rev| tr '[ATGC]' '[TACG]'"); else print $10}' file.sam

 

Categories: awk, UNIX