Tailer: A tool for 3' end analysis of non-polyadenylated RNAs from 3' sequencing data
Requirements
Python >= 3.6
pysam
Bio
gffuitls
requests
tqdm
Installation
pip install jla-tailer
Usage
Global alignment with a GTF annotation and SAM/BAM formatted files
Tailer -a [GTF Annotation] [SAM or BAM Files]
Required Arguments
-
-a, --annotation
- GTF formatted database to be used to infer 3' ends. Ensembl GTFs have worked well for this pipeline
-
[SAM or BAM Files]
- Space separated list of sam/bam formatted file locations to perform 3' end tail analysis
Local alignment with with FASTA/Q and specific Ensembl IDs of interest or reference fasta
Tailer -e [comma separated list of EnsIDs no spaces] [FASTA/Q files]
Tailer -f fasta_reference_file.fasta [FASTA/Q files]
Required Arguments
-
-e, --ensids
- Comma separated list of ensembl IDs. Either -e or -f inputs are required.
-
-f, --fasta
- FASTA formatted file to align reads against. Either -f or -e inputs are required
-
[Trimmed FASTQ files]
- Space separated FASTQ file locations for analysis.
Optional arguments
-
-t, --threshold [int, default=100]
- Any alignment identified further than this distance in nucleotides from the mature end will be considered spurious and discarded
-
-x, --trim [int, default=0]
- Helper for local mode only, can remove X nucleotides from adapter on the 3' end
-
-read, --read [int, default=1]
- Paired end only. 1 or 2 to signify which end contains the 3' end information.
-
-r, --rev_comp
- If set, will reverse complement the reads. Necessary for some library prep methods
-
-f, --fasta
- Use a fasta file as a reference instead of building one from ensembl IDs (Local Only)
-
-s, --sequence
- Output sequence in the tail file. Useful for debugging.
miRNA tailing (in development)
Tailer --miRNA [sam files]
Determines tails using an alignment file (SAM/BAM) that was aligned to a fasta genome of mature miRNAs
Optional arguments
-
--rpad [int, default=0]
- Number of downstream nucleotides added to distinguish between post-transcriptional and genome-encoded tails