Closed DrFengchenzhao closed 1 year ago
Hi @DrFengchenzhao ,
Thanks for the issue.
We sequenced bulk WGS, scRNA-seq and ST on the same sample. I wondered how I can harmonize these omics, especially using SNPs genotyped by bulk WGS to further increase power to detect CNVs in scRNA-seq.
Yes, this is possible. You can first run allele pileup using cellsnp-lite
with the WGS heterozygous SNP VCF. Then you run eagle2
phasing on the WGS VCF, and merge the phased GT fields with the allele counts to produce an allele dataframe in the format of Numbat input. Please refer to the underlying code in pileup_and_phase.R
script to see how to modify the procedure to do this.
Also, does 'via multi-sample mode of pileup-and-phase' mean that I should run pileup-and-phase.R with scRNA-seq and ST together?
Yes, you can do that. This will help increase allele coverage for co-genotyping.
Hey Teng,
I'm a bit lost after running cellsnp on the het regions, how do proceed after that? Would I attempt to the df_allele matrix from the cellsnp output?
Thanks, Chang
Hi @cnk113 ,
You can probably directly use the preprocess_allele
function to get df_allele
from the cell-snp counts and WGS phased VCF:
Hi Teng Gao,
I have implemented your methods on my scRNA-seq data which showed great advantages over others. I just noticed that you mentioned 'Therefore, a priori genotyping by DNA or co-genotyping with scRNA-seq (via multi-sample mode of pileup-and-phase) would be especially useful.' on suggestions of preparing spatial transcriptomics data.
We sequenced bulk WGS, scRNA-seq and ST on the same sample. I wondered how I can harmonize these omics, especially using SNPs genotyped by bulk WGS to further increase power to detect CNVs in scRNA-seq.
Also, does 'via multi-sample mode of pileup-and-phase' mean that I should run pileup-and-phase.R with scRNA-seq and ST together?
Thank you for your patience. Looking forward to your reply.
Dr. Chenzhao Feng