Thanls a lot I am goind to take a look at this ressource
Good morning,
I want to analyse public SHort Read archive from an RNA-seq experiment. I am trying to quantify them using sailfish. it is needed to provide the library type as an argument : -l "<libtype>" But I only that my data are paired-end ( dataset GSE89063) How can I know what to write for this argument ? Thanks a lot
Have a good day
3 answers
Hi, You can also check by performing some steps to know the strand specificity, please check the link below.
http://onetipperday.sterding.com/2012/07/how-to-tell-which-library-type-to-use.html
Hope it works with you.
Excellent resource to read
Unstranded
Kits:
TruSeq RNA Sample Prep kit
Parameters:
- HISAT2 / TopHat / Cufflinks / Cuffdiff: library-type fr-unstranded
- HTSeq: stranded -- no
Directional, first strand:
Kits
- All dUTP methods, NSR, NNSR
- TruSeq Stranded Total RNA Sample
- Prep Kit TruSeq Stranded mRNA Sample Prep Kit
- NEB Ultra Directional
- RNA Library Prep Kit Agilent SureSelect Strand-Specific
Parameters:
- HISAT2 / TopHat / Cufflinks / Cuffdiff: library-type fr-firststrand
- HTSeq: stranded -- reverse
Directional, second strand:
Kits:
- Directional Illumina (Ligation), Standard SOLiD
- ScriptSeq v2 RNA-Seq Library Preparation Kit
- SMARTer Stranded Total RNA
- Encore Complete RNA-Seq Library Systems
- NuGEN SoLo
Parameters:
- HISAT2 / TopHat / Cufflinks / Cuffdiff: library-type fr-secondstrand
- HTSeq: stranded -- yes
So first go to SRA and check which protocol they have used, whether strand specific or not. If strand specific, then which strand, that you can know by the kit they use.
Hope this helps
For that, we follow the approach AIR (https://transcriptomics.sequentiabiotech.com) uses. You should need a BAM file and an annotation file (BED file with the exon coordinates and the corresponding strand). Afterwards, you can use:
http://rseqc.sourceforge.net/#infer-experiment-py
To know whether the data is stranded or unstranded.
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Is there a reason you use Sailfish and not Salmon, which is more recent, and has an option to detect the library type?
I already tried salmon, I wanted to compare the results between Salmon and Sailfish