Handling Multiple Sequencing Runs in scRNA-seq
0
0
Entering edit mode
2.1 years ago
dk0319 ▴ 70

I am interested in analyzing some scRNA-seq data from GEO and I found that the samples have multiple runs (4 in some cases). I am wondering if someone has experience with this and if there is a recommended standard procedure for handling multiple runs?

For ChIP or bulk-rna seq I would just merge the files but I am unsure if this is normal for single-cell data.

scRNA-seq • 1.3k views
ADD COMMENT
1
Entering edit mode

Please check if the four sample named in pattern L001/001/003/004 which can be understood by sequencing was done in 4 lane of the sequencer and you can merge the fastq file to run your downstream analysis.

ADD REPLY
0
Entering edit mode

This is how the metadata is presented on SRA explorer. Just to confirm I should be able to merge these as 1?

"SRX15246019: GSM4274678: BMET1-Tumor; Homo sapiens; RNA-Seq 4 ILLUMINA (Illumina HiSeq 4000) runs: 130.2M spots, 12G bases, 5.3Gb downloads"

Runs: 4 runs, 130.2M spots, 12G bases, 5.3Gb"
Run # of Spots  # of Bases  Size    Published
SRR19180790 32,510,181  3G  1.3Gb   2022-05-17
SRR19180791 31,813,145  2.9G    1.3Gb   2022-05-17
SRR19180792 32,963,341  3G  1.3Gb   2022-05-17
SRR19180793 32,918,198  3G  1.3Gb   2022-05-17
ADD REPLY
0
Entering edit mode

It says it's RNA-seq.

ADD REPLY
0
Entering edit mode

Yes, but the data is scRNA-seq with R1, R2 and L1 fastq files

ADD REPLY
0
Entering edit mode

You are correct this is how the runs are labeled as L001-004. My question now is because each of these samples consists of R1, R2 and L1 fastq files, am I correct in assuming I need to merge all the R1s and R2s and L1s individually to produce 3 final fastq files?

ADD REPLY
0
Entering edit mode

Do you mean the samples come from different batches? In that case, you want to account for batch effect. You can find a nice explanation on how to account for batch effect here.

ADD REPLY

Login before adding your answer.

Traffic: 2600 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6