Single cell RNASeq data with only R2.fastq
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3.4 years ago

Hi everyone,

I downloaded some single cell RNA-Seq data from SRA sites. Unfortunately, only the R2.reads (~100bps) was uploaded for all the SRR files (SRR11573310...).

I am wondering if anyone would know if it will be ok to align the reads to genome just like regular RNA-Seq pipeline. For example, use hisat2_htseq-count to achieve the gene counts?

Thank you very much for any the help!

RNA-Seq • 937 views
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If this is 10X and R2 is only the cDNA then that would give you kind of "bulk" sample with excessive PCR duplicates since both the Cellular Barcodes (the per-cell identifier) as well as the UMIs (for deduplication) are in R1. I would contact the authors, scRNA-seq is complex enough, no need to add additional uncertainty by custom (and unreliable) pipelines.

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Thank you. I get the point. Would you help me understand why duplcates is a bigger problem for single cell RNASeq? I remember we generally don't remove duplicates for regular RNASeq. Many thanks!

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Read 1 may be available from Data Access tab for these accessions (an example here), if the name of the file is correct. Look under "Original Format".

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Thanks. I think they just name it as R1 but its actually R2 as I only can found one file. It is a big headaache to download single cell RNASeq data.

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