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In case of Drop-seq, we have paired end data.
Read 1: Cell code + UMI (unique molecule identifier)
Read 2: The transcript information
But I have a problem/doubt with the sample I am working on.
The sample I am using is the following:
(Check the "Reads" tab)
As you know the Drop-seq is "paired-end", we are expected to see two reads for a spot. Although this sample say paired-end, it has only one read per spot.
For example I can share a link of a different scRNA-seq data where you can properly see two reads for a spot
Example sample:
https://trace.ncbi.nlm.nih.gov/Trace...run=SRR8086553 (Check the "Reads" tab)
Where I am going wrong?
I asked one of the main authors of the paper. The following is the reply I got :
"I recommend that you download the aligned BAM files that are hosted in the same GEO record. Read 1 is already processed into the cell and UMI barcodes and held as custom tags (XC and XM) in the BAM files. The cells are already barcode-corrected, so if you use those files, your cell barcodes will line up with mine; if you start from FASTQs, they will not. For most aligners, you can just use the BAM file as input to realign. (It has all reads, even those that did not align.)"
But I could not find any "XM" or "XC" keywords in the bam file
To understand his reply you have to be familiar the processing steps of the Drop-seq: Link: https://github.com/broadinstitute/Dr...1.2Jan2016.pdf
Looks like they have submitted some kind of processed data. I could not figure out how much the data is processed. I am trying to use the data starting at different processing steps but I could not figure out how much the data is processed.
Read 1: Cell code + UMI (unique molecule identifier)
Read 2: The transcript information
But I have a problem/doubt with the sample I am working on.
The sample I am using is the following:
(Check the "Reads" tab)
As you know the Drop-seq is "paired-end", we are expected to see two reads for a spot. Although this sample say paired-end, it has only one read per spot.
For example I can share a link of a different scRNA-seq data where you can properly see two reads for a spot
Example sample:
https://trace.ncbi.nlm.nih.gov/Trace...run=SRR8086553 (Check the "Reads" tab)
Where I am going wrong?
I asked one of the main authors of the paper. The following is the reply I got :
"I recommend that you download the aligned BAM files that are hosted in the same GEO record. Read 1 is already processed into the cell and UMI barcodes and held as custom tags (XC and XM) in the BAM files. The cells are already barcode-corrected, so if you use those files, your cell barcodes will line up with mine; if you start from FASTQs, they will not. For most aligners, you can just use the BAM file as input to realign. (It has all reads, even those that did not align.)"
But I could not find any "XM" or "XC" keywords in the bam file
To understand his reply you have to be familiar the processing steps of the Drop-seq: Link: https://github.com/broadinstitute/Dr...1.2Jan2016.pdf
Looks like they have submitted some kind of processed data. I could not figure out how much the data is processed. I am trying to use the data starting at different processing steps but I could not figure out how much the data is processed.
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