We had some protein microarrays done I believe using the hu-prot platform. I asked for non-normalised corrected data hoping to run in DeSeq2 (as I have used this for RNAeq data before).
The count data imported as "cts" looks something like this:
RF0064 | RF0065 | RF0070 | |
COL4A3BP | 2927 | 4186 | 5818 |
IRAK4 | 9072 | 63923 | 5558 |
Patient ID's in columns and proteins in rows, obviously allot more of each. One note here I had to round values up so there where no decimal places for DeSeq
My metadata looks like this in "coldata":
id | case_control | days | card |
RF0064 | case | 33 | carditis |
RF0065 | control | 35 | control_NA |
RF0070 | case | 33 | non_carditis |
Again allot more patients.
Here is an example of case vs control analysis I did
dds_case_control <- DESeqDataSetFromMatrix(countData = cts,
colData = coldata,
design = ~ case_control)
dds_case_control$case_control <- relevel(dds_case_control$case_control, ref="control")
dds_case_control <- DESeq(dds_case_control)
res_case_control <- results(dds_case_control)
summary(res_case_control)
Which all seems to work fine, but I am not an expert at all with this and wondered if there is some ways within Deseq2 to quality control the results? Or if perhaps if I should use a different R package? I do like Deseq2 as I am now familiar with it, but ay advice welcome.
Thank you,
I assumed if I had some kind of data that was equivalent to “counts” I.e background subtracted and corrected data, so essentially have protein abundance data, that the analysis would be equivalent. Showing my ignorance I guess.
No, a fluescent intensity is not a count, and becomes even less like a count after background subtraction.