#!/bin/bash ################################################################################## #Andy Rampersaud, 02.01.16 #Adapted from tophat/paired_end_mapping scripts by Tisha Melia ################################################################################## #Assumptions for this job to run correctly: #1. This script is intended to map paired end RNA-Seq reads #2. You are mapping to mm9 (ucsc) reference genome #3. Your data is organized in the following way: #You have a data set dir such as: #wax-es/aramp10/G83_Samples #Within this dir you have sample specific folders such as: #G83_M1 #G83_M2 #G83_M3 #G83_M4 #Within each sample specific folder you have a "fastq" folder with: #File: *.fastq.gz such as: #G110_M1.fastq.gz ################################################################################## #Fill in the following information: ################################################################################## #Information about your data set #As mentioned above, you should have a data set dir containing your sample specific folders: #Dataset_DIR=/projectnb/wax-es/aramp10/G110_Samples ################################################################################## #Samples to process #To facilitate processing of samples in parallel we can use a text file that lists the samples to analyze #Note: this text file is still valid even if there is only one sample to process #You need to have a "Sample_Labels" dir within your Dataset_DIR #Within the Sample_Labels dir have a Sample_Labels.txt such that: ################################################ #The text file is formatted like the following: #---------------------------------------------- #Sample_DIR Sample_ID Description #Sample_Waxman-TP17 G83_M1 Male 8wk-pool 1 #Sample_Waxman-TP18 G83_M2 Male 8wk-pool 2 #Sample_Waxman-TP19 G83_M3 Female 8wk-pool 1 #Sample_Waxman-TP20 G83_M4 Female 8wk-pool 2 #---------------------------------------------- #The 1st column: The Sample_DIR name #The 2nd column: Waxman Lab Sample_ID #The 3rd column: Sample's description ################################################ #Sample_Labels_DIR=${Dataset_DIR}/Sample_Labels ################################################################################## #Bowtie2Index_DIR #Need this dir for the Bowtie2 Index #Feel free to use my dir but it's better practice to have a copy in your own Dataset_DIR #Bowtie2Index_DIR=/projectnb/wax-es/aramp10/Bowtie2 ################################################################################## #GTF_Files_DIR #Need this dir that contains the various GTF files #Feel free to use my dir but it's better practice to have a copy in your own Dataset_DIR #GTF_Files_DIR=/projectnb/wax-es/aramp10/GTF_Files ################################################################################## #STRANDEDNESS #There are three types of library construction. Please select your type of library construction # #1.) fr-unstranded (Standard Illumina) -> Reads from the left-most end of the fragment (in transcript coordinates) map to the transcript strand, and the right-most end maps to the opposite strand. #2.) fr-firststrand (dUTP, NSR, NNSR) -> Same as above except we enforce the rule that the right-most end of the fragment (in transcript coordinates) is the first sequenced (or only sequenced for single-end reads). Equivalently, it is assumed that only the strand generated during first strand synthesis is sequenced. #3.) fr-secondstrand (Ligation, Standard SOLiD) -> Same as above except we enforce the rule that the left-most end of the fragment (in transcript coordinates) is the first sequenced (or only sequenced for single-end reads). Equivalently, it is assumed that only the strand generated during second strand synthesis is sequenced. # # NOTE: If your dataset comes from Spira lab, your library type is likely to be "fr-unstranded". # If your dataset comes from MIT/Encode, your library type is likely to be "fr-firststrand". #If RNA-Seq library was prepared in-house using the NEBNext Ultra Directional kit, use option “fr-firststrand” #If your data is unstranded then use "fr-unstranded" #--------------------------------------------------------------------------------- #Choose one: #STRANDEDNESS="fr-unstranded" #STRANDEDNESS="fr-firststrand" #STRANDEDNESS="fr-secondstrand" #--------------------------------------------------------------------------------- ################################################################################## ################################################################################## #Please: DO NOT EDIT CODE BELOW ################################################################################## ################################################################################## #Gene Annotation file to use #--------------------------------------------------------------------------------- #For read mapping purposes we want the full gene body information (use the RefSeq_GeneBody.gtf) #Presence of splice junctions should be based on the full gene structure for any isoform of a gene symbol #For read counting purposes we can use either #1. RefSeq_GeneBody.gtf #2. Intron_Only_Regions.gtf #3. Exon_Only_Regions.gtf #--------------------------------------------------------------------------------- #ANNOTATION_FILE=RefSeq_GeneBody.gtf #--------------------------------------------------------------------------------- ################################################################################## #Time hour limit #On SCC a 12-hour runtime limit is enforced on all jobs, unless specified explicitly. #A runtime limit can be specified in the format "hh:mm:ss" #Dont change the following time limit value unless you know that your job is going to go over 12 hrs #TIME_LIMIT="12:00:00" ##################################################################################