Hello,
I hope all is well. So I am planning to run the ROSMAP samples using a transcriptome alignment tool, called Kallisto and I was wondering if you know specifically how the libraries were generated and in what direction sequencing was performed along the strand? I was looking at this paper by Bennet et al., 2018 (https://www.nature.com/articles/sdata2018142) and they mentioned the sequencing was done using the Illumina's HiSeq 2000 platform. I've analyzed data using Illumina's HiSeq 4000 before, and the core we sent our samples to sequenced them on the reverse strand (sense) first. Also I looked at more information on Synapse of how the libraries were generated (https://www.synapse.org/#!Synapse:syn23650893), but there wasn't any specifics of how the strand-specific library generation was performed.
So I am assuming, whether the ROSMAP samples were sequenced the same way on the reverse strand first using an Illumina HiSeq platform? If so, this information would help me as I perform transcriptome alignment next. If you can please clarify this information for me, then that would be really helpful.
Thank you again,
Phoebe
Created by Phoebe Valdes prvaldes Hi @prvaldes,
You are correct, the Rosmap libraries are reverse-stranded. I've tested this by aligning random samples using the STAR aligner and viewing the resulting gene counts as described in the star manual (https://github.com/alexdobin/STAR/blob/master/doc/STARmanual.pdf). I'm not sure specifically which Illumina HiSeq platform was used, but I haven't found any odd adapter sequences that would suggest that the specific platform used will influence your analyses.
Best,
Will
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